opus® 5, 10 - 60 cm columns opus® 45r, opus® 60r · pdf fileopus® 45r,...

OPUS® 5, 10 - 60 cm Columns OPUS® 45R, OPUS® 60R Columns Pre-packed Columns REGULATORY SUPPORT FILE

The information contained in this document is subject to change without notice. Repligen Corporation makes no warranty of any kind with regard to this material, including, but not limited to, the implied warranties of merchantability and fitness for a particular purpose. Repligen Corporation shall not be liable for errors contained herein or for incidental or consequential damages in connection with the furnishing, performance, or use of this material. No part of this document may be photocopied, reproduced, or translated to another language without the prior written consent of Repligen Corporation. For further information, please contact Repligen Corporation at www.repligen.com. © 2017 Repligen Corporation. All rights reserved. The trademarks mentioned herein are the property of Repligen Corporation and/or its affiliate(s) or their respective owners.

Customer Support [email protected] +1-781-250-0111 (option 2) Technical Support +1-781-250-0111 (option 3) [email protected] Repligen Corporation 41 Seyon Street Building #1, Suite 100 Waltham, MA 02453

www.repligen.com

mailto:[email protected]


http://www.repligen.com/

Contents

1. Introduction ......................................................................................................................... 6 2. Product Information ............................................................................................................. 7

2.1 Product Overview ...................................................................................................................... 7

2.2 Product Design ........................................................................................................................... 7

2.3 Certificate of Analysis (CoA) Examples ...................................................................................... 9

2.3.1 Pre-GMP OPUS® Columns Certificate of Analysis (CoA) ................................................. 9

2.3.2 GMP Run Ready OPUS® Columns Certificate of Analysis (CoA) .................................... 10

2.3.3 Certificate of Analysis: Chromatograms ....................................................................... 11

2.4 Column Manufacturing and Packing ........................................................................................ 13

2.5 Materials of Construction: Product Contact ............................................................................ 17

2.6 OPUS® Physical Specifications ................................................................................................. 22

2.7 Column Stability ....................................................................................................................... 23

2.7.1 Warranty ....................................................................................................................... 26

2.8 Material Safety Data Sheet ...................................................................................................... 26

3. Manufacturing Information ................................................................................................ 27 3.1 Introduction ............................................................................................................................. 27

3.2 Manufacturing Quality Assurance Standards and Policy ......................................................... 27

3.3 Manufacturing Facilities........................................................................................................... 27

3.4 Manufacturing Controls and SOPs ........................................................................................... 27

3.5 OPUS® Column Manufacturing ................................................................................................ 29

3.6 Bioburden and Endotoxin Release Testing .............................................................................. 34

4. Shipping Qualification ......................................................................................................... 35 4.1 Summary .................................................................................................................................. 35

4.2 Introduction ............................................................................................................................. 35

4.3 Method: ISTA 2A and 2B Tests ................................................................................................. 37

4.3.1 ISTA Test Methods ........................................................................................................ 37

4.3.2 Column Efficiency Test Methods .................................................................................. 39

4.3.3 Results: ISTA and Column Efficiency Tests .................................................................... 39

4.3.4 Chromatographic Performance Test Results ................................................................ 40

4.3.5 Conclusion ..................................................................................................................... 40

5. Extractables and Leachables ............................................................................................... 41 5.1 Introduction and Background .................................................................................................. 41

5.2 OPUS® Extractables Strategy ................................................................................................... 42

5.3 USP<88> Biological Reactivity Tests, In Vivo............................................................................ 42

5.3.1 CFR 21 Part 177 Indirect food Additives: Polymers ...................................................... 43

5.3.2 USP General Chapters 661: Polypropylene Containers ............................................... 43

5.3.3 Animal Free and EMEA 410/01 ..................................................................................... 43

5.4 Extractable Test Protocol and Results ..................................................................................... 44

5.4.1 Extruded PP: OPUS® 10 – 30 cm Columns Only ............................................................ 46

5.4.2 Machined Compression Molded PP: OPUS® 10 – 60/R cm Columns ............................ 47

5.4.3 PP Mesh: OPUS® 10 – 60/R cm Columns ...................................................................... 48

5.4.4 P/ E-Glass Composite: OPUS® 45/R – 60/R cm Columns Only ...................................... 49

5.4.5 Platinum Cured Silicone O-Ring: OPUS® 10 – 60/R cm Columns .................................. 50

5.4.6 Platinum Cured Silicone Braided Tubing: OPUS® 10 – 60/R cm Columns .................... 50

5.4.7 Conclusions ................................................................................................................... 51

5.5 Leachables: OPUS® Column Strategy ....................................................................................... 52

5.6 Considerations for Leachables Testing .................................................................................... 52

5.7 References ............................................................................................................................... 53

6. Appendix ............................................................................................................................ 54 6.1 OPUS® 5, 10-60/R cm ID Material Certificates (available upon request) ................................ 54

6.2 Engineering Drawings .............................................................................................................. 55

6.2.1 OPUS® 5 cm Column ..................................................................................................... 55

6.2.2 OPUS® 10 cm Column ................................................................................................... 56

6.2.3 OPUS® 14 cm ID Column ............................................................................................... 57

6.2.4 OPUS® 20 cm Column ................................................................................................... 58

6.2.5 OPUS® 25 cm Column ................................................................................................... 59

6.2.6 OPUS® 30 cm Column ................................................................................................... 60

6.2.7 OPUS® 45 cm and 45R Column ..................................................................................... 61

6.2.8 OPUS® 60 cm and 60R Column ..................................................................................... 62

6.3 Packaging ................................................................................................................................. 63

6.3.1 OPUS® 5 cm Column Packaging .................................................................................... 63

6.3.2 OPUS® 10 - 14 cm Column Packaging ........................................................................... 64

6.3.3 OPUS® 20 - 30 cm Column Packaging ........................................................................... 65

6.3.4 OPUS® 45 cm Column Packaging .................................................................................. 66

6.3.5 OPUS® 60 cm Column Packaging .................................................................................. 67

List of Tables

Table 2.1 OPUS® Pre-GMP and GMP Applications ............................................................................... 8 Table 2.2 Product Contact Materials .................................................................................................. 21 Table 2.3 Physical Specifications Summary on OPUS® 10 - 30 cm Columns ...................................... 22 Table 2.4 Physical Specifications Summary on OPUS® 45R - 60R Columns ....................................... 22 Table 2.5 Column Mass (Approximate Weight of Packed Columns).................................................. 23 Table 2.6 Stability Study Schedule and Acceptance Criteria .............................................................. 23 Table 2.7 Mechanical Stability Test Results ....................................................................................... 24 Table 2.8 Performance Stability Study Results .................................................................................. 24 Table 2.9 Results of Bioburden and Endotoxin Testing ..................................................................... 25 Table 3.1 Repligen Water Specifications Compared with ASTM, USP Purified Water and Water

for Injection ........................................................................................................................ 29 Table 3.2 Parts Cleaning Specifications .............................................................................................. 32 Table 4.1 ISTA 2A and 2B Testing Conditions (OPUS® 5, 10 – 60 cm Columns) ............................... 37 Table 4.2 OPUS® 10 cm, 20 cm and 45 cm Column Performance Data ............................................ 40 Table 5.1 Summary of USP <88> Plastics Classes .............................................................................. 42 Table 5.2 Polymer List and Regulatory Information .......................................................................... 45

List of Figures

Figure 2.1 Pre-GMP OPUS® Column Certificate of Analysis ................................................................ 9 Figure 2.2 GMP Run Ready OPUS® Column Certificate of Analysis................................................... 10 Figure 2.3 OPUS® Certificate of Analysis Testing Conditions and Chromatograms .......................... 11 Figure 2.4 Example of OPUS® 10-60 cm ID Column Label ................................................................. 12 Figure 2.5 OPUS® Part Number Format ............................................................................................ 12 Figure 2.6 Lifting Handles and Input/Output Ports on an OPUS® 20 cm Column ............................. 16 Figure 2.7 Castor Detail on an OPUS® 45 cm Column ....................................................................... 16 Figure 2.8 Materials of Construction on OPUS® 5 cm Column ......................................................... 17 Figure 2.9 Materials of Construction on OPUS® 10-30 cm Columns................................................. 18 Figure 2.10 Materials of Construction on OPUS® 45-60 cm Columns ................................................ 18 Figure 2.11 Materials of Construction on OPUS® 45R - 60R Columns ................................................ 19 Figure 2.12 OPUS® R Port Design and Materials of Construction ....................................................... 19 Figure 3.1 OPUS Work Order Example (blank) .................................................................................. 30 Figure 4.1 OPUS® 5 cm Column Shipping Container Design ............................................................. 35 Figure 4.2 OPUS® 10 - 30 cm Column Shipping Container Design .................................................... 36 Figure 4.3 OPUS® 45/R - 60/R Column Chipping Container Design .................................................. 36 Figure 4.4 OPUS® 5 cm Column Compression and Drop Testing ...................................................... 38 Figure 4.5 OPUS® 20 cm Column Vibrational and Compression Testing .......................................... 38 Figure 4.6 OPUS® 60/R Column Vibrational, Incline Impact and Drop Testing ................................. 39

6

OPUS® 5, 10 – 60 cm, OPUS® R Columns Regulatory Support File

1. Introduction

The Regulatory Support File (RSF) for OPUS® (Open platform, User-specified) Pre-Packed Columns is intended to be used as:

A guide for appropriate application use in process development, clinical, and commercial

purification processes

A guide to validation in manufacturing processes

A support reference for CMC submissions for regulatory license approval

A guide for supplier audits

In place of a Drug Master File submission

This regulatory support file covers all column IDs including OPUS® 5, 10, 14, 20, 25, 30, 45, 60, 45R, 60R Columns. Unless otherwise specified all information applicable to OPUS® 45 and 60 designs is also applicable to OPUS® 45R and 60R designs. Repligen is committed to providing all relevant technical, manufacturing, and quality information, however, only non-confidential information is presented in this document. Confidential details may be made available upon request through a formal confidentiality agreement or as part of a supplier audit. Repligen’s Quality Policy

Copies of Repligen’s quality policy, quality manual, and ISO certificate can be found on the following

website, https://www.repligen.com/resources/quality-documents/.

Safety Notices

Follow all local regulations for safe disposal

For laboratory and manufacturing production only

Responsible Official The individual designated responsible for quality and regulatory affairs for Repligen, and to whom all correspondence or requests for audits should be addressed.

Senior Director of Quality

Telephone: +1-781.250.0111

Email: [email protected]

https://www.repligen.com/resources/quality-documents/


7


2. Product Information

2.1 Product Overview OPUS® Pre-packed Columns (Open platform, User-specified ) columns are designed to perform chromatographic purification of biological molecules as part of the production process in either GMP or non-GMP applications. The OPUS® Pre-packed Column platform offers an alternative to traditional self-pack glass columns, and can be reliably packed with nearly any bioprocessing resin. To accommodate a wide range of biopharmaceutical applications, OPUS® columns are configurable for bed height and available in a range of industry standard internal diameters.

2.2 Product Design

The OPUS® 5, 10 – 60/R cm Column platform is designed to meet the requirements of GMP manufacturing in the pharmaceutical and biopharmaceutical industries for campaign-use applications.

Platformable OPUS® Columns are configurable to accommodate a wide range of purification applications for vaccines, monoclonal antibodies, and recombinant proteins. For example, an end user can configure an OPUS® Column by choosing a diameter and specifying:

Column bed height

Chromatography resin

Repligen recommended release criteria, or customized release criteria (ex: plate count, asymmetry, storage solution, etc.)

Quality Standards In order to meet the needs of GMP manufacturing, OPUS® 5, and 10 – 60/R cm ID columns are manufactured in the U.S.A. under the following quality standards:

Repligen maintains an ISO 9001 compliant Quality Management System that is currently certified

by BSI Americas. A copy of the current ISO certification can be downloaded from the following

web address, https://www.repligen.com/resources/quality-documents/.

All materials in direct fluid contact path meet USP Class VI, and USP <88> requirements for In Vivo

Biological Reactivity

All columns are packed in a controlled, classified clean room that meets ISO Class 7 Non-Viable

Particulate (NVP) standards

All fluid contact components are free from materials of animal origin or compliant with EMEA

410/01 Rev 3

In order to provide the most flexibility to end users, OPUS® Pre-Packed columns are available in two levels for non-GMP applications and GMP applications. See Table 2.1 for more detail. In general, OPUS® Pre-GMP and GMP Run Ready columns are identical except as noted in Table 2.1. GMP Run Ready columns come with a resin side sample for incoming resin identity testing at an end-users facility. OPUS® Pre-GMP columns do not come with a resin side sample.


8


Table 2.1 OPUS® Pre-GMP and GMP Applications

Certificate of Analysis Component Pre-GMP

10 – 60/R cm ID’s GMP Run Ready

5, 10 – 60/R cm ID’s

GMP Level X X

Catalog Number X X

Lot Number X X

Serial Number X X

Column Diameter X X

Final/Achieved Bed Height X X

Chromatography Resin Type X X

Chromatography Resin Lot Number X X

Shipping/Storage Solution X X

Date of Manufacture X X

Customer Property Number X X

Asymmetry Specification and Result X X

Theoretical plate count (N/m) Specification and Result

X X

Chromatography Release Test Chromatogram X X

ISO 9001 Compliance Statement X X

Product Contact Materials Compliance Statement (animal free, EMEA 410/01, USP Class VI)

X X

Packing Environment Statement X X

Chromatography Resin Control Statement X

Copy of Resin Certificate of Analysis X X

Endotoxin Specification and Test Result (pass/fail)

X

Bioburden Specification Test Result (pass/fail) X

9


2.3 Certificate of Analysis (CoA) Examples

Each OPUS® Pre-packed Column is delivered with a Certificate of Analysis (CoA) with the content noted in Table 2.1.

2.3.1 Pre-GMP OPUS® Columns Certificate of Analysis (CoA)

Each Pre-GMP column produced is issued a Certificates of Analysis (CoA). Content is derived from the template information (Figure 2.1) as well as customer required specifications as detailed on the OPUS® Work Order. The CoA package also includes the resin manufacturer’s CoA which is attached to the OPUS® Column CoA.

Figure 2.1 Pre-GMP OPUS® Column Certificate of Analysis

10


2.3.2 GMP Run Ready OPUS® Columns Certificate of Analysis (CoA)

Each GMP Run Ready column produced is issued a Certificates of Analysis (CoA). Content is derived from the template information (Figure 2.2) as well as customer required specifications as detailed on the OPUS® Work Order. The CoA package also includes the resin manufacturer’s CoA which is attached to the OPUS® Column CoA. In addition, all GMP Run Ready columns are provided with a chromatography resin side sample which is shipped with the column and resin lot traceability.

Figure 2.2 GMP Run Ready OPUS® Column Certificate of Analysis

11


2.3.3 Certificate of Analysis: Chromatograms

For both Pre-GMP, and GMP Run Ready columns, the Certificate of Analysis comes with a second page detailing the testing conditions and chromatogram. See example below of the second page of the OPUS® GMP Run Ready CoA. Figure 2.3 OPUS® Certificate of Analysis Testing Conditions and Chromatograms

Labeling Each individual OPUS® 10 – 60/R cm ID Column is labeled with the following information:

1. Catalog number

2. Resin type

3. Column Internal Diameter

4. Column Bed Height

5. Date of Manufacture

6. Storage temperature (“ambient” or “2-8 degrees Celsius”)

7. Lot number

8. Serial number

9. Pressure Rating

12


Figure 2.4 Example of OPUS® 10-60 cm ID Column Label

Each individual 5cm ID GMP Run Ready column is labeled with the following 1. Catalog Number 2. Resin description 3. Lot number 4. Serial number

OPUS® Column Part Numbers OPUS® Column part numbers are structured so basic column configuration details can be deciphered. The OPUS® part number system, which is governed by QA-FM-10107, is not unique for all user specified inputs, and therefore an OPUS® Work Order is generated for every new purchase order even if the same column/part number is re-ordered. See Figure 2.5 for the OPUS® part number format. Figure 2.5 OPUS® Part Number Format

13


If for example a customer were to order a 45 x 15 cm GMP Run Ready OPUS® R Column packed with Repligen-sourced CaptivA® resin, the part number would be, BC-450-CPRI-150-GR. The part number code works as follows:

The first 2 digits (“BC”) show the resin is Repligen sourced (vs. customer supplied)

The next 3 digits show the nominal column diameter in millimeters (“450”)

The subsequent 4 digits show the unique code for resin being packed (“CPRI”)

The next 3 digits show the target/specified bed height in millimeters (“150”),

The last 1 or 2 digits reference the GMP level (Pre-GMP or GMP Run Ready) as well as if the column configuration includes an OPUS® R port (“GR”)

2.4 Column Manufacturing and Packing

Obtaining User Specified Column Inputs

All OPUS® columns are manufactured using input from the OPUS® quote request form (https://www.repligen.com/rfq/), which documents user specified configurable product aspects such as,

Column details: internal diameter, bed height, resin source, GMP Level

Acceptance criteria (if specified): plate count, asymmetry, pressure vs. flow

Application specific (if specified): maximum working flow rate, maximum working pressure,

preferred storage solution

All fields on the OPUS® Quote Request form are filled in with input from the end user and/or a Repligen sales professional. The form is then submitted for review prior to the formal quoting process. Once a purchase order has been submitted and accepted against a formal quotation, an OPUS® Work Order is drafted by Repligen to formalize the user specified inputs contained in the quote request form. The Work Order is then sent to the customer for review and approval. Upon written approval from the customer, the OPUS® Work Order (QA-FM-10016) is finalized and the column/s is added to the manufacturing schedule. See Section 3 for more information on the OPUS® Work Order.

Resin Supply

When ordering an OPUS® column, the resin may be supplied by the customer, drop shipped from the resin manufacturer, or procured through Repligen. If the customer decides to source the resin and supply it to Repligen for column packing, then the Health and Safety Declaration for Accepting Customer Supplied Resin form (QA-FM-02-10035) must be completed. Overview of OPUS® Column Production Processes* Column Components: All incoming raw materials and custom components are subject to an incoming inspection procedure. Only the parts in compliance with the criteria set forward in the approved Raw Materials and Component Specifications documents are released for column assembly and packing. Resins: For all resins, Repligen maintains strict material controls including assignment of internal lot numbers to enable full traceability within batch records consistent with good documentation practices. The SOPs which govern these procedures are SOP-1094 (Control of Incoming Materials),

14


SOP-10042 (Control of Incoming Components) and SOP-10035 (Customer Property). In addition, all resins are tested for identity (FTIR) prior to release for column packing.

Manufacturing Qualification: The column assembly processes are qualified and documented with appropriate SOPs and batch records as specified in Repligen’s ISO 9001 Quality Management System.

Preparation of Column Components: Column parts are washed and cleaned prior to assembly. Washing and cleaning procedures are qualified and validated.

Column Packing: OPUS® 5, 10 – 60/R cm ID columns are packed in a controlled, classified clean room that meets ISO Class 7 standards.

Quality Control: Each pre-packed column is individually tested according to the specifications detailed on the OPUS® Work Order. The specifications and the results of the release tests are documented on the Certificate of Analysis.

Packaging: Columns are secured by foam packaging material, and shipped in qualified shipping containers.

The OPUS® 5, and 10 – 30 cm columns are sealed in polyolefin bags in a controlled, classified clean room that meets ISO Class 7 NVP standards and then packaged in a cardboard box with foam packaging material. For OPUS® 10 - 30 cm columns, the box is strapped and shipped on a plastic pallet. See Sections 4 and 6 for more information.

The OPUS® 45R – 60R cm columns are not bagged; however, the inlet and outlet ports are both capped and secured with SaniSure clamps. All OPUS® 45R – 60R cm columns are shipped in a custom made wooden crate. All crates are heat treated and stamped with heat treated assurance. OPUS® 45 and 45R columns ship in the same qualified crate design, and OPUS® 60 and 60R ship in the same qualified crate design. See Sections 4 and 6 for more information.

The box/crate and internal packaging for the OPUS® 5, and 10 - 60/R cm ID columns have been qualified with standard International Safe Transit Authority (ISTA) tests. For more information on shipping qualification, please refer to Section 4 for Shipping Qualifications.

*Note: See Section 3 for more detailed information on the OPUS® Column production process. Reference Documentation Quality Documentation: Industry standards for document control are followed as per Repligen ISO 9001 Quality Management System. Repligen Quality management documents can be found on the following website, https://www.repligen.com/resources/quality-documents/. Technical Specifications: External column dimensions are provided in the user guide and in Section 2.5 (Table 2.3 and 2.4) of this Regulatory Support File. All other column packing specifications and user specified criteria are documented in the OPUS® Work Order. User Guide: A user guide for OPUS® 5, 10-60/R columns is available on Repligen’s website, as well as an instruction guide for unpacking resin using the OPUS® R design. Please visit https://www.repligen.com/resources/quality-documents/.

Un-packaging Guide: For OPUS®45-60/R columns, an uncrating guide is available on Repligen’s website. Please visit, https://www.repligen.com/resources/quality-documents/.




15


Technical Documents: A variety of supporting technical documents, presentations, and videos can be found on Repligen’s website by visiting www.repligen.com/opus. CE-Certificate: EG directive 97/23/EG is not applicable for OPUS® columns, and therefore no columns will be CE-signed.

Performance OPUS® columns are designed to provide chromatographic performance equivalent to or better than traditional self-packed columns. The performance is attained through a semi-automated packing process and the design of the flow distribution system.

Optimal flow distribution is achieved through a flow distributor design with minimized dead

space volume. Rigorous Computational Fluid Dynamics (CFD) models validated by experiments

have provided evidence this flow distributor design is optimal for uniform radial distribution.

Mathematical modeling and performance testing have verified that the flow distributor design

provides chromatographic performance consistent with traditional columns.

Multi-cycle performance tests have shown that OPUS® columns deliver robust and reproducible

chromatographic results

Note: More information on column performance can be found on at www.repligen.com/opus. Ease of Use The OPUS® 5, 10 – 60/R cm ID platform design incorporates ease of use features such as lifting

handles, castors, and industry standard sanitary tri-clamp inlet and outlet connections (see Table for

more information).

Inlet/Outlet Port Locations The OPUS® 10-60/R column inlet and outlet ports are both located on the top side of the column.

The column inlet and outlet are secured with a tri-clamp fitting designed by SaniSure® to be both

leak-proof and tamper-evident.

The OPUS® 5 cm column design has inlet and outlet ports located at either end of the tube like a typical lab scale self-pack column design.

http://www.repligen.com/opus

http://www.repligen.com/opus

16


Lifting Handles

OPUS® columns with 14 – 30 cm IDs are designed with handles as shown below. The OPUS® 5, 10,

45/R, and 60/R Columns have no lifting handles. Columns with internal diameters of 5 cm through

30 cm may be lifted safely by one or two operators.

Figure 2.6 Lifting Handles and Input/Output Ports on an OPUS® 20 cm Column

Castors

OPUS® 45/R – 60/R cm ID columns are equipped with twin wheel swivel castors shown below for

enhanced maneuverability. Each castor has a manually engaged wheel lock and is rated to hold

100kg. The bottom caps for OPUS® 45/R and 60/R Columns are designed with grooves to allow

lifting of the columns with straps.

Figure 2.7 Castor Detail on an OPUS® 45 cm Column

17


2.5 Materials of Construction: Product Contact OPUS® 5 and 10 - 60/R cm ID columns are designed using plastics which are best suited for downstream processing applications. These plastics have been chosen for performance characteristics including: Fit for Purpose

Suitability for pharmaceutical use with process fluid contact

USP Class VI compatibility

Low levels of non-toxic extractables

Free from materials of animal origin and/or compliant with EMEA 410/01

Mechanical Strength Recommended maximum 4 bar operating pressure for 5, and 10 – 30 cm IDs, and maximum 3 bar operating pressure for 45/R – 60/R cm ID columns. All columns were designed with a ≥ 2x Factor of Safety (FOS) in operating pressure.

Thermal Stability: Column construction can support a working temperature range of 2°C to 40°C based on the general properties of polypropylene as well as OPUS® Column ISTA shipping tests (see Section 4). Figure 2.8 Materials of Construction: OPUS® 5 cm Column

18


Figure 2.9 Materials of Construction: OPUS® 10-30 cm Columns

Figure 2.10 Materials of Construction: OPUS® 45-60 cm Columns

19


Figure 2.11 Materials of Construction: OPUS® 45R - 60R Columns

Figure 2.12 OPUS® R Port Design and Materials of Construction

20


Materials of Construction: Product Contact

OPUS® 5, and 10 – 60/R cm ID columns are made of polymeric materials chosen to provide optimum biological and chemical compatibility with a vast range of biopharmaceutical manufacturing strategies. The elegant design of 10 – 60/R cm ID OPUS® columns employ only two product contact materials, polypropylene and platinum cured silicone. The 5 cm ID columns employ only polypropylene. All plastics have been certified to comply with United States Pharmacopeia (USP) Class VI requirements as defined in USP <88> Biological Reactivity Tests and USP <661> Physicochemical Tests, In Vivo as well as FDA CFR Title 21 Part 177. Furthermore all product contact plastics are animal free and/or compliant with EMEA 410/01 guidance.

Product Contact Materials Summary

OPUS® 5, and 10 – 30 cm ID Columns

o The same medical grade polypropylene homo-polymer formulation is used for the

column tube, flow distributors, inlet, outlet, and bed support screens

o The polypropylene bed support screens are made from a medical grade

polypropylene formulation

Note: the polypropylene bed support screen is ultrasonically welded to the

flow distributor.

o The flow distributor O-rings are made from platinum-cured medical grade silicone

o The return line (10-30cm columns only) is made from platinum-cured, medical grade

silicone, reinforced with a strong polyester fiber

OPUS® 45/R – 60/R cm Columns

o Column tubes are made from a 70% w/w E-glass/Polypropylene engineered

composite structure assuring a polypropylene resin rich internal surface. This

material exhibits a high strength-to-weight ratio, excellent toughness, and chemical

resistance.

o Flow distributors, inlet ports, outlet ports, OPUS® R plug, and OPUS® R inside port

are made from the same medical grade polypropylene homo-polymer formulation

o The polypropylene bed support screens are made from a medical grade

polypropylene formulation

Note: the polypropylene bed support screen is ultrasonically welded to the

flow distributor.

o The flow distributor O-rings, OPUS® R plug O-ring, and the inner/outer OPUS® R

gaskets are made from the same platinum-cured, medical grade silicone

o The return line is made from platinum-cured medical grade silicone, reinforced with

a strong polyester fiber

21


Table 2.2 Product Contact Materials

Component Material USP <88>

Class

CFR 21 177 Animal

Origin

Column Tube (5,

10-30 cm)

Polypropylene Class VI 177.1520 Animal Free

Column Tube (45/R

– 60/R)

70% w/w E-Glass / PP engineered

composite structure

Class VI 177.1520 Animal Free

Flow Distributors Polypropylene Class VI 177.1520 Animal Free

Inlet and Outlet

Ports


Bed Support

Screens

Polypropylene Class VI 177.1520 EMEA

410/01

Flow Distributor O-

Rings

Platinum-cured medical grade silicone Class VI 177.2600 Animal Free

Return Line Platinum-cured medical grade silicone,

reinforced with strong polyester fiber

Class VI 177.2600 Animal Free

OPUS® R Plug, and

Inside Port


OPUS® R Inner/

Outer Gaskets, and

Plug O-ring

Platinum-cured medical grade silicone Class VI 177.2600 Animal Free

Non-Product Contact Materials Summary

Top and bottom caps: Acrylonitrile Butadiene Styrene (ABS) co-polymer or High Density

Polyethylene (HDPE)

Side guard: ABS (blue)

Inlet and outlet port grommets: silicone (blue)

Handles: polyurethane (blue)

Castors: Polyamide casing with non-marking grey polyurethane tires

OPUS® R outer nut and washer: ABS

OPUS® R BioClamp: glass filled nylon

22


2.6 OPUS® Physical Specifications

Table 2.3 Physical Specifications Summary on OPUS® 10 - 30 cm Columns

Column Diameter

Physical Attributes

5 cm 10 cm 14 cm 20 cm 25 cm 30 cm

Internal cross section

20.4 cm2 78.5 cm

2 154 cm

2 314 cm

2 491 cm

2 707 cm

2

Column Body Pressure Rating

4 Bar 4 Bar 4 Bar 4 Bar 4 Bar 4 Bar

Bed height range 5 – 30 cm 5 – 30 cm 5 – 30 cm 5 – 30 cm 5 – 30 cm 5 – 30 cm

Column Volumes

10 cm bed height

20 cm bed height

30 cm bed height

0.2L

0.4L

0.6L

0.8L

1.6L

2.4L

1.5 L

3.1 L

4.6 L

3.1 L

6.3 L

9.4 L

4.9 L

9.8 L

14.7 L

7.1 L

14.1 L

21.2 L

Assembled Column Height (cm)

~20 + bed height

~20 + bed height

~30 + bed height

~30 + bed height

~33 + bed height

~35 + bed height

Outer Diameter (including caps)

6.3 cm 16 cm 21 cm 27 cm 33 cm 38 cm

Inlet/Outlet Flow Path Internal

Diameter

3.18 mm

0.125”

6.35 mm

0.25”

6.35 mm

0.25”

6.35 mm

0.25”

9.53 mm

0.375”

9.53 mm

0.375”

Inlet and Outlet Port Connectors

¾” mini tri-clamp






Table 2.4 Physical Specifications Summary on OPUS® 45R - 60R Columns

Column Diameter Column Diameter

Physical Attributes 45.7 cm (includes R design) 59.9 cm (includes R design)

Internal cross section 1,640 cm2 2,817 cm

2

Column Body

Pressure Rating 3 Bar 3 bar

Bed height range 5 – 30 cm 5 – 30 cm

10 cm bed height CV

20 cm bed height CV

30 cm bed height CV

16 L

33 L

49 L

28 L

56 L

84 L

Assembled Column Height (cm) ~ 90 cm ~92 cm

Outer Diameter (including caps) 54 cm 61 cm

Inlet/Outlet Flow Path Internal Diameter

10.88 mm

0.43 inches

19.05 mm

0.75 inches

Inlet and Outlet Port Connectors Per ASME BPE Standards, Current Edition

¾” mini tri-Clamp 1” tri-Clamp

23


Table 2.5 Column Mass (Approximate Weight of Packed Columns)

Column Diameter

Bed

Height 5 cm 10 cm 14 cm 20 cm 25 cm 30 cm 45 cm 60 cm

5 cm 1 kg 2 kg 4 kg 6 kg 10 kg 14 kg 69 kg 117 kg

10 cm 1.2 kg 2.5 kg 5 kg 8 kg 13 kg 18 kg 77 kg 131 kg

15 cm 1.4 kg 3 kg 6 kg 10 kg 16 kg 22 kg 86 kg 145 kg

20 cm 1.5 kg 3.5 kg 7 kg 12 kg 19 kg 26 kg 94 kg 159 kg

30 cm 1.9 kg 4 kg 9 kg 16 kg 24 kg 34 kg 110 kg 187 kg

2.7 Column Stability Repligen conducted a study to assess shelf life stability for OPUS® columns when stored according to instructions in the user guide. Two different studies ran in parallel over an 18 month period:

Mechanical stability study: This study looked at the structural integrity of the column assembly at pre-determined time points over the course of 18 months.

Study design: Three columns of different internal diameters (10 cm, 20 cm, and 30 cm ID) initially filled with 18.5% EtOH solution up to a 20 cm bed height were pressurized to 4 bar and the pressure decay was monitored for 60 minutes. During the course of this pressure hold period, the column was visually inspected for gross failure such as leakage from ports and/or threads, and general structural integrity. See Table 2.6 for the mechanical stability test schedule and acceptance criteria. Results are summarized in Table 2.7.

Chromatographic performance stability study: This study looked at the chromatographic performance of the large scale columns over an 18 month period when stored at 2-8 Celsius. The goal of this study was to show no significant performance degradation will occur if a customer purchases a column and places it on the shelf for up to 18 months. Cold storage was deemed as “worst case” due to the polypropylene materials becoming more brittle in cold temperatures.

Study Design: Four 20 cm columns were packed with agarose resin to a 15 cm bed height. Acceptance criteria for the columns is summarized in Table 2.6 at “T=0.” One column was retested at each of the time intervals and compared with the performance of that specific column at time zero. The results are summarized in Table 2.8.

Table 2.6 Stability Study Schedule and Acceptance Criteria

Time Interval Mechanical Stability Criteria

(Empty Columns)

Performance Stability Criteria

(Packed Columns)

T = 0 Time at Pressure: 60 minutes Pressure: 4 bar Pressure decay: < 10% Visual: No gross defects or leaks

Asymmetry: 0.8 – 1.6 Plate Count: >1500 N/m (100cm/hr) P vs. F: record baseline (10 psi) Endotoxin: ≤ 1.0 EU/mL Bioburden: ≤ 100 CFU/mL

24


T = 3 months Time at Pressure: 60 minutes Pressure: 4 bar Pressure decay: < 10% Visual: No gross defects or leaks








All columns passed acceptance criteria at each interim time point, as well as at the end of 18 months. The data is summarized in Tables 2.7 and 2.8 below:

Table 2.7 Mechanical Stability Test Results

Column Configuration

3 months 6 months 12 months 18 months

10 x 20 cm Pass Pass Pass Pass



Table 2.8 Performance Stability Study Results

Column S/N 3 months 6 months 12 months 18 months

0001 N/m Δ = < 15% As Δ = < 15%

N/m Δ = < 15% As Δ = < 15%

0002

N/m Δ = < 15% As Δ = < 15%

N/m Δ = < 15% As Δ = < 15%

0003

N/m Δ = < 15% As Δ = < 15%

N/m Δ = < 15% As Δ = < 15%

0004

N/m Δ = < 15% As Δ = < 15%

25


At the end of 18 months, bioburden for all 4 packed columns (serial numbers 0001 – 0004) was 0 CFU/ml and endotoxin for all 4 packed columns (serial numbers 0001-0004) was <0.1 EU/ml.

Cleanability

OPUS® Columns are used for purification of biological products which have specific regulatory

requirements for bioburden and endotoxin levels. Therefore, a quantitative cleaning investigation

was performed to demonstrate the effectiveness of sanitization using sodium hydroxide as a

cleaning agent.

Method: Several different OPUS® Columns were packed with Sepharose 6FF in 20 cm bed heights.

Each column was loaded with 1 column volume of E. coli bacteria at a concentration of 0.1 OD

(optical density) at 600 nm. The columns were left to incubate at ambient temperature overnight,

and then flushed with reverse osmosis deionized (RODI) water for 2-3 column volumes in down-

flow. Samples were collected to measure bioburden and endotoxin. A sanitization procedure was

subsequently used as followed:

Sanitization procedure: • Flush with 1 M sodium hydroxide in up-flow at 100 cm/h for 30 minutes

• Flush with 1 M sodium hydroxide in down-flow at 100 cm/h for 30 minutes

• Recirculation of 1 M sodium hydroxide for 2 hours in up-flow at 100 cm/h

• Incubation of the column in 1 M sodium hydroxide for 1 hour (static sanitization for compete

removal of endotoxins)

• Flush with RODI water at 100 cm/h until neutral pH is achieved

Each column was then left to incubate at ambient temperature overnight prior to pulling samples for

bioburden and endotoxin testing.

Results: Results for bioburden and endotoxin levels from the microbial challenge are outlined in the

table below, which shows the sanitization procedure completely removed bioburden from millions

of CFU to zero CFU in the post-sanitization water rinse. In addition, endotoxin levels were brought

below the limit of detection (0.25 EU/mL) for the assay.

Table 2.9 Results of Bioburden and Endotoxin Testing

Column Diameter

Cleaning Data 20 cm 30 cm 45 cm 45R cm 60 cm 60R

0.1 M NaCl

effluent after

overnight E.coli

incubation

TNTC (CFU/Ml)

>0.25 EU/mL

TNTC (CFU/Ml)

>0.25 EU/mL

TNTC (CFU/Ml)

>0.25 EU/mL

TNTC (CFU/Ml)

>0.25 EU/mL

TNTC (CFU/Ml)

>0.25 EU/mL

TNTC (CFU/Ml)

>0.25 EU/mL

0.1 M NaCl post

sanitization

effluent

0 CFU/mL

<0.25 EU/mL

0 CFU/mL

<0.25 EU/mL

0 CFU/mL

<0.25 EU/mL

0 CFU/mL

<0.25 EU/mL

0 CFU/mL

<0.25 EU/mL

0 CFU/mL

<0.25 EU/mL

26


Conclusions: In order to test effectiveness of sanitization on an OPUS® column, a worst case scenario was devised where the column was loaded with an excess of E. coli culture (a gram-negative, endotoxin producing bacteria). The results of the sanitization protocol demonstrate the effective removal of bioburden and endotoxin contamination.

2.7.1 Warranty

Repligen aspires to complete customer satisfaction, and has implemented the following warranty policy for OPUS® Columns.

If the column arrives at the customer site in damaged condition, Repligen will accept the damage risk and issue a replacement at no charge.

If the column fails passing specifications as agreed to on the OPUS® Work Order, Repligen will conduct troubleshooting efforts and if unsuccessful, will ask for the column to be returned for further evaluation. If the root cause of the failed test is determined to be a compromised packed bed, Repligen will pack a replacement at no charge.

If the column’s packed bed is compromised (i.e. a channel develops) and the root cause is packing, then Repligen will replace the column at no charge.

2.8 Material Safety Data Sheet OPUS® columns are made from plastic components only and therefore no MSDS is needed. An MSDS for the chromatography resin packed into an OPUS® Column may be obtained from the resin manufacturer.

27


3. Manufacturing Information

3.1 Introduction All OPUS® 5, 10 – 60/R cm Columns are manufactured at the Repligen corporate headquarters, located at 41 Seyon Street, Waltham, Massachusetts 02453, USA. The QA and QC operations are also based in the same location. Neither the facility nor products manufactured require registration or market approval. Therefore, Repligen’s column packing facility and products manufactured herein are not subject to regulatory review or audit by organizations such as the US Food and Drug Administration or European Medicines Agency.

3.2 Manufacturing Quality Assurance Standards and Policy Repligen recognizes the need for high quality standards, and has therefore established an ISO 9001 Quality Management System suitable for the needs of our clients and our industry. Refer to Section 1 for Repligen’s Quality Policy.

3.3 Manufacturing Facilities The OPUS® Column packing suite consists of multiple ISO Class 7 column packing rooms and a central ISO 8 prep area. Two airlock doorways are maintained, one for people and one for finished goods and equipment. In general, column parts enter the suite through a validated parts washer or are cleaned with NaOH followed by a rinse with RODI water. The OPUS® Column suite is completely segregated from all other product manufacturing at Repligen and is a restricted-access area. The clean-room environment is controlled and monitored as follows:

Air quality is maintained by 100% HEPA filtered air

Preparation room air quality is tested to ISO Class 8 standards for non-viable particulates

Column packing room air quality is tested to ISO Class 7 standards for non-viable particulates

Room pressure differentials are maintained and monitored according to SOPs

All rooms are on a routine cleaning and disinfection schedule

Access is restricted to authorized personnel only

Gowning is required for entry into controlled areas including a secondary gowning procedure for

entry into the ISO Class 7 packing rooms

Environmental monitoring is performed to check for viable and non-viable contamination

3.4 Manufacturing Controls and SOPs

Training [SOP-1498]: Manufacturing is performed by qualified and trained operators. Training documentation is maintained by Quality Assurance. Process Documentation [SOP-1302, SOP-1098]: Repligen manufacturing processes are governed by an ISO 9001 compliant Quality Management System. All manufacturing work instructions are contained in controlled documents, which are issued in advance of each manufacturing batch. Batches and process intermediates are 100% traceable through an internal lot numbering system. All manufacturing data are recorded by operators at the time of manufacturing. Batch records are archived for 3 years on site, and then stored off site for a minimum of 10 years.

28


Raw Materials [SOP-1094]: All raw materials are controlled, and each raw material has a pre-approved specification. Receipt of material is verified and released by QA prior to use in manufacturing. Customer Property [SOP-10035]: All customer property including customer supplied resin is documented, tracked, and released prior to the start of packing. Customer property is stored in segregated areas within Repligen’s warehouse. Supplier Management [SOP-1496]: Repligen identifies critical suppliers of raw materials and components based on the impact to the quality of the product they are used to manufacture. Critical suppliers are subject to a qualification process and are monitored and routinely audited according to a pre-determined schedule. The supplier audit schedule is established based on critical supplier audit cycle, supplier performance, past audit results, and business requirements. Components [SOP-10042]: All components are controlled and each component has a pre-approved specification. Every receipt of material is verified and released by QA prior to use in manufacturing. In addition, each OPUS® Column component has a pre-approved engineering drawing, and is subject to incoming QC inspection, which includes review of materials certificates, and verification of critical dimensions. Process Change Management [SOP-1447]: Manufacturing process changes are governed by change management procedures that include provisions for customer notification of major changes. Product Storage Control [SOP-MFG-1212; SOP-1263]: Product is stored either at ambient temperature, or in temperature controlled units as dictated by the packed resin. All temperature controlled storage units have backup power, 24/7 electronic monitoring, and alarms that are monitored. Preventive Maintenance [SOP-1501] and Calibration [SOP-1181]: Equipment and monitoring devices are controlled through the Repligen Equipment Control process [SOP1171]. Each piece of equipment is uniquely identified and has a preventive maintenance and/or calibration schedule as necessary. High Purity Water [SOP-MFG-1209]: Purified water is supplied to all manufacturing areas from a Reverse Osmosis/Deionization (RODI) system. The RODI system is fully automated, and provides high quality water in a continuously circulating loop. The water system design performance specifications are ASTM Type I Reagent Grade Water, with the addition of low endotoxin and bioburden specifications (Table 4.1). Water quality is monitored continuously using an on-line TOC device, and is routinely sampled and tested by Repligen Quality Control.

29


Table 3.1 Repligen Water Specifications Compared with ASTM, USP Purified Water and Water for Injection

Parameter ASTM Type I USP Purified

water

WFI Repligen

Specification

Conductivity 0.05 µS/cm 1.3 µS/cm 1.3 µS/cm < 0.01 mS/cm

Endotoxin N/A N/A < 0.25 EU/mL < 0.5 EU/mL

Bioburden N/A 100 CFU/mL 0.1 CFU/mL ≤ 10 CFU/mL

pH N/A N/A 5 – 7 5 – 7

TOC 0.1 ppm 0.5 ppm 0.5 ppm 0.1 ppm

High purity water is one of the key raw materials for any bioprocessing product. Repligen believes that these specifications, in conjunction with regular maintenance, ensure that the water system performs at a suitably high quality standard.

3.5 OPUS® Column Manufacturing

Packing Environment and Environmental Controls All OPUS® columns are packed in ISO Class 7 classified clean rooms and buffers prepared in an ISO 8 classified clean room. The rooms are monitored on a weekly basis for viable contaminants using TSA settle plates. Microbial contaminants (bacteria) are counted and reported. Viable contamination limits have been set for the ISO 8 rooms as follows:

Alert Limit: > 20 CFU/plate Action Limit: > 35 CFU/plate

Viable contamination limits have been set for the ISO 7 rooms as follows: Alert Limit: > 10 CFU/plate Action Limit: > 25 CFU/plate Counts which exceed an alert or action limit are investigated according to SOP-MFG-1207.

The OPUS® manufacturing suite is monitored on a weekly basis for Non-Viable Particles (≥ 0.5 µm). Each room is monitored in predetermined locations, measuring 1 cubic foot of air over the course of 1 minute. NVP limits have been set for the ISO Class 8 clean room spaces as follows:

Alert Limit: ≥ 25,000 particles per ft3 (880,000 particles per m3) Action Limit: ≥ 50,000 particles per ft3 (1,760,000 particles per m3) ISO 8 Specification: ≤ 100,000 particles per ft3 (3,520,000 particles per m3)

NVP limits have been set for the ISO Class 7 clean room spaces as follows: Alert Limit: ≥ 2,500 particles per ft3 (88,000 particles per m3) Action Limit: ≥ 5,000 particles per ft3 (176,000 particles per m3) ISO 7 Specification: ≤ 10,000 particles per ft3 (352,000 particles per m3)

30


OPUS® Work Order and User Specified Inputs Once a purchase order for a column has been received, the OPUS® Work Order (QA-FM-10015) is created. The OPUS® Work Order compiles information from the purchase order as well as the user specified inputs which were recorded from the OPUS® quote request form (https://www.repligen.com/rfq/). Note: All OPUS® columns are built to order and each column ordered must have a quote request form and unique Work Order. These forms document critical user specified inputs and therefore cannot be eliminated from the ordering process. In addition, each OPUS® Work Order is unique to a PO number and column part number. The OPUS® Work Order captures a variety of information including the following,

Quote number, purchase order number and column part number

Column size, type, and resin

Column packing method and solutions used

Column performance release method and specifications (ex: Asymmetry and plate count)

Revision history Once all required fields have been populated, Repligen sends the OPUS® Work Order to the appropriate customer contact for review. If all the details in the document are satisfactory, the customer signs and dates the form, and then sends the signed form back to Repligen. Once the signed form is received, Repligen can begin column packing. Note: Column packing cannot begin without a customer approved work order. See below for an example of the OPUS® Work Order. Figure 3.1 OPUS Work Order Example (blank)

https://www.repligen.com/rfq/

31


Chromatography Resin Control All chromatography resins are tested for identity using FTIR prior to being released for production. A copy of the FTIR spectrum is maintained with the completed batch record. The SOPs governing chromatography resin control procedures are SOP-1094 (Control of Incoming Materials), SOP-10042 (Control of Incoming Components) and SOP-10035 (Customer Property).

Repligen Supplied Resins Resins purchased and supplied by Repligen for column packing are controlled as a raw material according to SOP-1094 (Control of Incoming Materials). Upon arrival, each resin is placed into quarantine pending disposition by Repligen Quality assurance. Upon release, each batch of resin is given a unique internal lot number and expiration date to ensure full traceability throughout the manufacturing process. Customer Supplied Resins Repligen’s customer property procedures (SOP-10035) are designed to protect our customer’s financial and intellectual property interests to prevent unauthorized use, disclosure, or disposal. All resins shipped to Repligen for column packing must be unused. Each customer supplied resin is assigned a unique customer property number which is typically linked to a specific purchase order and part number. The customer property number is noted on the form entitled, Health and Safety Declaration for Accepting Customer Supplied Resin, which must be filled out by an end user prior to shipping resin to Repligen. Upon arrival, the resin is managed according to SOP-10035, which dictates the resin must be labeled, segregated, and then stored according to the resin vendor’s recommendation. All customer supplied resin is also tested for identity using FTIR prior to being released for production. Unused media is either returned or discarded according to the customer’s instructions.

Repligen understands in some instances customers may have a requirement to sample and test a small aliquot of the resin for identity (ex. FTIR). Unused resin in containers, which have been sampled for QC testing, are compatible with Repligen’s quality requirements and therefore can be accepted for column packing.

Resin Preparation Once the appropriate resin has been released, parts cleaned, and column packing batch record documentation have been issued, the follow steps occur: 1. Manufacturing operators move sealed resin containers into one of the OPUS® ISO 7 clean

rooms. Note: only the released resin is allowed in the packing room, and only one resin type at a time can be present.

2. Manufacturing operators verify the appropriate resin by checking against the OPUS® Work Order.

3. After verifying the resin, containers are opened for resin preparation (decantation of storage solution, fines removal, etc.).

4. For OPUS® GMP Run Ready columns only, prior to beginning resin prep an operator removes 10-30 mL of resin per resin lot from the original container. This operation is performed using a sterile pipette and the operator dispenses the resin aliquot into a 60 mL gamma irradiated, tamper evident PETG (polyethylene terephthalate) bottle. The bottle is then sealed and labeled with resin name, lot number, part number, date, and Repligen operator initials. In the event multiple resin lots are packed into a single column, a single side sample of each resin lot will be taken. Therefore, multiple side samples will be delivered with a GMP Run Ready column if multiple resin lots are packed.

32


5. After column packing, the resin “QC sample/s” or “resin side sample/s” are stored with the finished the OPUS® GMP Run Ready column for shipment.

In general, Repligen uses the following approach for slurry preparation and percent solids determination: 1. Decant off storage solution 2. Perform defining of resin as specified by resin MFG. 3. Suspended resin in packing buffer to target the desired % solids 4. Combine resin bottles in a holding tank if required 5. Pull resin samples 6. Record mass and volume added for density calculations 7. Spin samples in a centrifuge 8. Read spin down solids volume 9. Determine average % solids (solids vol. / total vol.) 10. Use target compression factor (CF = target column vol./uncompressed resin vol.) as derived by

a. Past empirical data b. Resin manufacturer recommendations c. Repligen R&D recommendations

11. Calculate target solids volume to add (target bed volume / CF) 12. Apply % solids (Step 9) and density (Step 6) to target solids calculations (Step 11) to determine

mass of slurry required 13. (Target solids vol. / % solids) * density = mass slurry to dose Assembly, Packing, and Performance Testing OPUS® columns are assembled from qualified and controlled components. Inspected and released component parts are taken from inventory and brought into the ISO 8 classified cleanroom through an automated parts washer. Parts which do not enter through the parts washer are chemically cleaned with NaOH, followed by rinse with RODI water to depyrogenate and reduce bioburden. Both processes have been qualified and validated to the following specifications:

Table 3.2 Parts Cleaning Specifications

Method Parts Cleaning Specifications

Bioburden Endotoxin TOC Conductivity pH

Parts Washer < 10 cfu/ml < 0.5 EU/ml < 10 PPM < 10 uS/cm 5.0 – 7.0

NaOH Soak < 10 cfu/ml < 0.25 EU/ml N/A < 50 uS/cm 5.0 - 7.0

The following points summarize the major steps which occur during OPUS® Column packing: 1. Solutions and buffers are prepared in the ISO 8 area of the OPUS® clean rooms. All buffers and

solutions are filtered with a 0.2um filter. 2. Packing solutions/buffers and allocated parts, which have been cleaned, are brought into the

designated ISO 7 packing room. 3. All subsequent steps are performed in the designated ISO 7 clean room. 4. The bottom flow distributor with the ultrasonically welded mesh is inserted into the bottom end

of the tube to form a water-tight seal. The flow distributor assembly is then secured in place with a polypropylene weld or adhesive bond.

33


5. The chromatography resin is added to the open ended column by dosing directly into the column or using a peristaltic pump.

6. The top flow distributor is inserted into the column tube and packing is initiated to establish a uniform bed. OPUS® columns utilize flow packing, axial compression, and/or a combination of both methods to achieve a well packed bed. In general, packing protocols are developed in Repligen’s R&D department and subsequently transferred to the production team.

7. Following the prescribed packing procedure, the top flow distributor is precisely positioned to create the correct resin compression. Note: Rigid particle resins such as ceramic hydroxyapatite are not compressed, rather the top flow distributor is positioned so there is no significant gap between the consolidated resin bed and the flow distributor assembly.

8. Bed performance is tested according to the procedure and specifications as documented on the OPUS® Work Order. Note: A column may be tested and flow distributor adjusted during the packing process to ensure optimal performance.

9. Once the final flow distributor position is set and the column has met the performance specifications (theoretical plates/meter and Asymmetry) as documented on the OPUS® Work Order, the packing process can conclude.

10. The column is then sanitized using a procedure recommended by the resin manufacturer and/or as documented on the OPUS® Work Order.

11. Post sanitization, the column is then neutralized (typically with RODI water), and Bioburden or Endotoxin samples taken (OPUS® GMP Run Ready Columns only).

12. After neutralization, each pre-packed column is put in its final storage solution as documented on the OPUS® Work Order.

13. The column assembly is then finished by securing the flow distributors in place using a polypropylene weld or adhesive bond, and assembling the top cap, bottom cap, and side-guard. Note: The top cap, bottom cap, and side guard serve cosmetic and ergonomic functions, and therefore do not impact the performance of the column.

14. Inlet and outlet ports are capped and sealed using a tamper evident SaniSure clamp. 15. Each OPUS® 5, 10 – 30 cm Column is sealed in a polyethylene bag before being removed from

the OPUS® cleanroom suite. Note: OPUS® 45/R and 60/R cm Columns are not bagged since they are on wheels.

16. Columns are then stored according to storage conditions as noted on the OPUS® Work Order.

Client resin verification upon arrival: Included with each column lot is a copy of the resin vendor’s Certificate of Analysis1 and a small sample/s (10-30 mL) of resin (GMP Run Ready columns only) intended for incoming quality control analysis. As described above, in the event multiple resin lots are packed into a column, a resin aliquot is taken for each resin lot and delivered with the column. Finally, the column label contains the resin description, and the certificate of analysis for each column indicates the part number and lot number of the pre-packed resin.

Equipment Used in the Manufacturing of OPUS® Columns: Wherever possible, all equipment used to pack OPUS® columns is dedicated single use equipment such as:

Slurry tanks and carboys

Single-use liners for buffer tanks

Single-use platinum cured tubing for pipe transfers

1 In rare cases a resin vendor’s certificate of analysis may not be included with the OPUS CoA package. For

example, when a customer ships custom resin without a CoA to Repligen for packing, Repligen may not be able to include a resin CoA in the OPUS CoA package if the custom resin CoA is not readily available from the manufacturer.

34


Where reusable components are used in the manufacturing process which might come into contact with the resin or the inside of the column, Repligen employs validated cleaning protocols to ensure the risk of cross resin “contamination” is mitigated.

3.6 Bioburden and Endotoxin Release Testing Columns are tested according to the specifications detailed in the OPUS® Work Order. Bioburden and endotoxin testing is performed on OPUS® “GMP Run Ready” columns and samples are taken in RODI water post sanitization and just prior to introducing the storage solution. The release specifications are as follows:

Bioburden: <10 CFU/mL

Endotoxin: <0.25 EU/mL Repligen performs endotoxin and bioburden testing in-house using the following two methods. Bioburden assay method Bioburden is analyzed by a vacuum filtration method. The sample is applied to a sterile filter under vacuum and then rinsed with peptone water. The filter is collected aseptically and placed onto TSA media. The plate is placed into an incubator at 32C +/- 2C for 5 days. An analyst reads the plates at day 3 and day 5 and documents the results. A negative control of the peptone water is also plated and incubated to demonstrate the reagents used during the analysis were not contaminated. Endotoxin assay method Endotoxin is analyzed by gel clot method. Repligen uses a commercially available kit and performs the test according to the manufacturer’s instructions which have been transcribed into an internal test procedure. A column rinse sample with a volume of 0.25 mL is introduced to the gel clot tube and incubated in a 37C water bath. The tubes are then read by an analyst and the data is recorded. A positive control is run alongside the sample for this assay. Bioburden and Endotoxin test results are documented on the Certificate of Analysis as a pass/fail result for each GMP Run Ready column. See Section 2.0 for sample CoA documents. Note: OPUS® Pre-GMP columns are not tested for bioburden or endotoxin.

35


4. Shipping Qualification

4.1 Summary

To certify the packaging, column components, and column performance remains intact during and after shipping, International Safe Transit Association (ISTA) tests were conducted on a variety of OPUS® column sizes. A third party certified laboratory performed the shipping tests, and Repligen tested chromatographic performance pre and post shipping. Results from the studies show the packaging withstands the stressors of commercial shipping, and chromatographic performance is maintained.

4.2 Introduction Objective The objective of the shipping studies is to demonstrate the custom made packaging (Figures 5.1 and 5.2) and pre-packed columns of multiple diameters arrive intact after worst case shipping simulations. See Section 6 for engineering drawings of packaging used for each internal diameter column. OPUS® columns employ different packaging based on size and weight. To cover the range of different column dimensions and packaging methods/materials the following column sizes were evaluated.

OPUS® 5 cm Column

OPUS® 10 cm Column (uses identical packaging as the OPUS® 14 cm Column)

OPUS® 20 cm Column (uses identical packaging as the OPUS® 25, and 30 cm Columns)

OPUS® 45 (uses identical packaging as the OPUS® 45R Column)

OPUS® 60 (uses identical packaging as the OPUS® 60R Column)

OPUS® 60R (representative of the “R” design) Figure 4.1 OPUS® 5 cm Column Shipping Container Design

36


Figure 4.2 OPUS® 10 - 30 cm Column Shipping Container Design

Figure 4.3 OPUS® 45/R - 60/R Column Shipping Container Design

Acceptance Criteria The shipping tests documented in this regulatory support file have two different acceptance criteria categories as detailed below.

Visual inspection of packaging Per ISTA guidelines, procedures 2A and 2B call for a simple pass/fail assessment based on a visual test of packaging materials. This assessment is made by the certified ISTA test facility and documented in a summary report.

Chromatographic performance Repligen assessed chromatographic performance by measuring plate count and asymmetry to compare “pre-shipping” performance with “post-shipping” performance. In order to pass the chromatographic portion of the test, the minimum post-shipping theoretical plate count and asymmetry measurements had to be within +/-25% of pre-shipping results.

37


4.3 Method: ISTA 2A and 2B Tests

4.3.1 ISTA Test Methods

ISTA Procedure 2A was used for the OPUS® 5 cm ID and 2B2 was used for OPUS® 10 – 60/R cm IDs. The selection of the two tests has to do with weight and shipping methods (see below).

ISTA Procedure 2A is a worst case simulation test for individual packaged-products shipped in boxes (“parcel shipments).

ISTA Procedure 2B is a worst case simulation test for individual packaged-products shipped on pallets

Both tests are used to:

Evaluate the performance of a packaged product

Compare relative performance of the package and product design

During the test, the package and product are considered together and not separately.

Tests and Conditions All studies were performed by Atlas Box and Crate in Sutton, MA per ISTA guidelines. The specific test conditions are described further in Table 5.1 below. The 5, 10, 20, 45, 60, and 60R columns were manufactured by Repligen, packaged into their respective shipping containers, picked up by Atlas, shipped to their certified testing facility, and tested according the applicable standards. After the tests were complete, Atlas returned the shipping containers to Repligen for post testing analysis.

Table 4.1 ISTA 2A and 2B Testing Conditions (OPUS® 5, 10 – 60 cm Columns)

Test Name Test Details

Atmospheric Conditioning Ambient Conditions for 36 hours

Controlled Temperature and Humidity

Conditioning 38° ±2°C; Relative Humidity 85% ±5% for 36

hours

4°C±2°C; RH uncontrolled for 36 hours

Compression Testing 248 lbs. (OPUS® 5), 370 lbs. (OPUS® 10), 744 lbs (OPUS®

20), 1728 lbs (OPUS® 45), 2,392 lbs (OPUS® 60), and

2,836 lbs (OPUS® 60 R) on top of crate/box

Random Vibration Testing 60 minutes of random vibration

Frequency (Hz) PSD Level, g2/Hz

1.0 0.0001

4.0 0.01

100.0 0.01

200.0 0.001

Incline Impact (2B only) Slide crate down ramp and into a wall

Drop Testing Crate dropped 8” to ground (2B)

2 See, www.ista.org/forms/2Boverview.pdf. Accessed on 02/04/2014

38


Crate bottom edges dropped 8” to ground (2B)

Box dropped 38 inches to the ground (2A)

Random Vibration Testing 60 minutes of random vibration

Frequency (Hz) PSD Level, g2/Hz

1.0 0.0001

4.0 0.01

100.0 0.01

200.0 0.001

Figure 4.4 OPUS® 5 cm Column Compression and Drop Testing

Figure 4.5 OPUS® 20 cm Column Vibrational and Compression Testing

39


Figure 4.6 OPUS® 60/R Column Vibrational, Incline Impact and Drop Testing

Visual Inspection

Per ISTA Procedures 2A and 2B, the certified test facility performed a visual pass/fail inspection of the packaging materials.

4.3.2 Column Efficiency Test Methods

Overview: Column efficiency testing was designed to assess and uncover any discernible differences in chromatographic performance pre-shipping and post-shipping. Repligen conducted all chromatographic performance tests.

5 cm Study: 5 cm x 10 cm OPUS® Column packed with Sepharose® 6 Fast Flow

10 – 14 cm Study: 10 cm x 15.3 cm OPUS® Column packed with Q Sepharose® Fast Flow resin

20 – 30 cm Study: 20 cm x 19 cm OPUS® Column packed with a silica based protein A resin

45 cm Study: 45 cm x 21 cm OPUS® Column packed with Q Sepharose® Fast Flow resin

60 cm Study: 60 cm x 20 cm OPUS® Column packed with Sepharose® 6 Fast Flow resin

60R Study: 60 x 20.5 cm OPUS® Column packed with 90% Phenyl Sepharose 6 Fast Flow with 10% SP Sepharose XL

Each column was tested at 100 cm/hr with pulse injection of 1% CV acetone solution, except for the OPUS® 5 cm Column which was tested at 100 cm/hr using a 0.2M NaCl mobile phase and 2M NaCl spike.

4.3.3 Results: ISTA and Column Efficiency Tests

Results: Visual Inspection ISTA test facility:

After the worst case shipping simulation tests were completed, Atlas visually inspected the packaging for signs of damage. For all tests, the custom designed OPUS® packaging passed the visual inspection without any significant signs of damage. Note: Visual inspection results are documented as part of the ISTA summary reports issued by the test facility.

Repligen:

Upon receipt from Atlas Box and Crate, all shipping containers and columns were inspected and found to be undamaged and intact.

40


Note: visual inspection results are documented in Repligen’s summary report for each shipping test

4.3.4 Chromatographic Performance Test Results

The data summarized in Table 5.2 shows the packing efficiency of 10, 20, 45, 60, and 60R OPUS® columns remain stable following the applicable shipping simulation. Stability is exhibited by a negligible change in asymmetry and plate count, while meeting the acceptance criteria. Table 4.2 OPUS® 10 cm, 20 cm and 45 cm Column Performance Data

Column Date N/m As Pass / Fail

5 cm ID Pre-shipping 2479 1.5 PASS

Post-shipping 3108 1.5 PASS









60R Pre-shipping 3367 1.0 PASS

Post-shipping 3373 1.0 PASS *Pre-ship and post-ship test flow rates vary slightly due to manual flow control of testing set-up.

4.3.5 Conclusion

In summary, the following was observed post worst case shipping simulation:

No significant damage to the shipping containers

No damage to the column structures

No discernable damage to the packed beds

No significant differences in chromatographic performance

Therefore, OPUS® columns designed and tested by Repligen for chromatographic performance, and the shipping containers designed and tested in conjunction with Atlas Box and Crate, demonstrate suitable robustness for surviving the harsh environments of commercial shipping.

41


5. Extractables and Leachables

5.1 Introduction and Background

Plastic materials have been used in the manufacturing of therapeutics for many decades. Over the last 15 years, focused product development by many vendors and biotechnology companies has resulted in a plethora of single-use technologies. During this time, the industry has witnessed the development and adoption of critical disposable and single-use technologies like mixers, bioreactors, filters, and connectors. As a result, standards and best practices for evaluating component safety have been set for the selection and qualification of plastics.

In general, plastics used in biopharmaceutical manufacturing have low defined extractables and have been determined to be non-toxic at equivalent therapeutic doses. Many base standards used for guidance have been set by regulatory publications including USP, CFR 21, and EMEA. These basic standards have been elaborated on by industry organizations like the Bio-Process Systems Alliance (BPSA) and Parenteral Drug Association (PDA) as well as product manufacturers through the publication of best practices of testing and assessment of data. In addition, end user therapeutic manufacturers have become more demanding in their analytical requirements, assessment of data, determination of risk, and minimum threshold for meeting internal standards.

Repligen is sensitive to the demands of the industry and will therefore supply relevant and applicable information about the plastics used in the product contact components of OPUS® columns. In accordance with industry standards, Repligen uses the definitions for extractables and leachables as stated in the 2007 BPSA document, "Recommendations for Extractables and Leachables Testing: Part 1.”

Extractables: Chemical compounds that migrate from any product-contact material when exposed

to an appropriate solvent under exaggerated conditions of time and temperature.

Leachables: Chemical compounds, typically a subset of extractables that migrate into a drug

formulation from any product contact material as a result of direct contact under normal process

conditions.

There is a general consensus that it is the responsibility of the product technology vendor to provide an extractable data package. In recent years there have been efforts to standardize the testing procedures for extractables, and wherever possible it is Repligen’s intent to comply with the standardization efforts. In addition, in compliance with CFR 21, Part 211.65, OPUS® columns are designed such that all product contact materials are not reactive, additive, or absorptive so as to alter the safety, identity, strength, quality, or purity of the drug product beyond established requirements.

Therefore, using the BPSA guidance as a backdrop, Repligen has designed an extractables program to produce a robust data package by conducting extractables testing using three solvents as well as exaggerated time and temperature conditions. Solvents were chosen for their common usage in chromatography columns. Leachables, however, are considered to be process specific and the responsibility of the end user to define within specific process parameters.

42


5.2 OPUS® Extractables Strategy Based on the above discussion, Repligen has applied the following philosophy in its approach to developing a meaningful extractables and leachables package for OPUS® columns:

1. All plastics used will be certified to meet four criteria (see Sections 3.2.1–3.2.4 for more detail):

a) Certified to meet USP <88> Biological Reactivity Tests, In Vivo (USP Class VI) b) Certified to meet CFR 21 Part 177 Indirect food Additives: Polymers

i. Sec. 177.1520 Olefin polymers (Polypropylene) ii. Sec. 177.2600 Rubber articles intended for repeated use (Silicone)

c) USP General Chapters 661: Polypropylene Containers d) Certified Animal Free or compliant with EMEA 410/01

2. Extractables from the OPUS® 5, and 10 – 60/R cm Column platform were determined according

to an approved written experimental rationale for exaggerated time and temperature conditions

and tested against an approved protocol.

3. Leachables from the OPUS® 5, and 10 – 60/R cm Column platform: Repligen will not conduct,

nor present data for leachables in this Regulatory Support File for any process specific solution

beyond what is determined in the extractables testing. Repligen will on request, support client

based leachable testing by providing component test materials.

5.3 USP<88> Biological Reactivity Tests, In Vivo Six plastic classes are defined in Table 3.1. This classification is based on responses to a series of in vivo tests for which extracts, materials, and routes of administration are specified. These tests are directly related to the intended end-use of the plastic articles.

Table 5.1 Summary of USP <88> Plastics Classes

Plastic Classes Tests to be Conducted

I II III IV V VI Extract Species Procedure

X X X X X X USP 0.9% NaCl Mouse Systemic-Intravenous

X X X X X X Rabbit Intracutaneous Irritation

X X X X X 1:20 EtOH/NaCl Mouse Systemic-Intravenous

X X X X X Rabbit Intracutaneous Irritation

X X X PEG 400 Mouse Systemic-Intravenous

X X Rabbit Intracutaneous Irritation

X X X X Cottonseed Oil Mouse Systemic-Intravenous

X X X Rabbit Intracutaneous Irritation

X X Muscle Implant Rabbit Intramuscular

(7 day, no Histopathology)

43


Acute Systemic Toxicity Test Purpose: In-vivo systemic tests evaluate the impairment or activation of a system rather than the impairment of individual cells or organs. In “acute” systemic toxicity tests, the test material (extract) is tested for systemic toxic effects as a result of a single, acute exposure. This test is designed to evaluate systemic responses to the extracts of materials following injection into mice.

Irritation Test: Intracutaneous Irritation Test

Purpose: The irritation tests are in-vivo screening tests to evaluate the potential of test materials or their extracts to cause irritation on the exposed part of the body. This test is designed to evaluate local responses to the extracts of materials following intracutaneous injection into rabbits.

Implantation Test: Intramuscular Purpose: Implant studies evaluate the local pathological effects on living tissue at both the gross and microscopic level of a test article surgically implanted into an appropriate implant site. The implantation test is designed for the evaluation of plastic materials and other polymeric materials in direct contact with living tissue.

5.3.1 CFR 21 Part 177 Indirect food Additives: Polymers

Subpart B: Substances for Use as Basic Components of Single and Repeated Use Food Contact Surfaces. Section 177.1520 – Olefin Polymers (Applied to Polypropylene). The olefin polymers from polypropylene referenced in CFR 21 Part 177 Sec. 177.1520 may be safely used as articles or components of articles intended for use in contact with food, subject to the provisions of this section. Subpart C: Substances for use only as components of articles intended for repeated use Sec. 177.2600, and rubber articles intended for repeated use (applied to silicone). Rubber articles intended for repeated use may be safely used in producing, manufacturing, packing, processing, preparing, treating, packaging, transporting, or holding food, subject to the provisions of this section.

5.3.2 USP General Chapters 661: Polypropylene Containers

The standards and test provided in this section characterize polypropylene containers produced from homopolymers or co-polymers that are suitable for packaging dry solid or liquid oral dosage forms.

5.3.3 Animal Free and EMEA 410/01

Note: This is for guidance on minimizing the risk of transmitting animal spongiform encephalopathy agents via human and veterinary medicinal products. Scientific principles for minimizing risk (EMEA 410/01): Use of materials from ‘non TSE-relevant animal species’ or non-animal origin is preferred. If materials from ‘TSE-relevant animal species’ have to be used, consideration should be given to all the necessary measures to minimize the risk of transmission of TSE.

44


5.4 Extractable Test Protocol and Results

By Michael Ruberto, Ph.D.

Background Repligen has performed polymer deformulation and extractables testing on the product contact components of their OPUS® columns. This data has been consolidated into a format that will be suitable for use in the OPUS® product literature. Extractables and Leachables Materials The materials used to fabricate single-use processing equipment for biopharmaceutical manufacturing are often polymers, such as plastic or elastomers (rubber), as opposed to the traditional metal or glass. Polymers offer more versatility, since they are light-weight, flexible and much more durable than their traditional counterparts. Plastic and rubber are also disposable, so issues associated with cleaning and its validation are often avoidable. Whenever there is contact between a material of construction for a manufacturing component and therapeutic, there is always the possibility for chemicals from the material to migrate or leach into the drug. All materials can produce leachables. Leachables are the chemicals that migrate from single-use processing equipment into the various components of the drug product during manufacturing. Extractables are chemicals entities, organic and inorganic, that can be extracted from single-use processing equipment using common laboratory solvents in controlled experiments. They represent the worst case scenario and are used as a tool to predict the types of leachables that can be encountered during pharmaceutical production. So, extractables are the “potentials” and leachables are the “actuals.” An effective way to minimize and control leachables is to use well characterized materials to fabricate the manufacturing equipment. The OPUS® column product contact materials are mainly constructed from polypropylene (PP) homopolymers and platinum cured silicone. PP homopolymer is much less prone to leachables than other types of polyolefins due to its mechanical and physical properties (such as melting point, glass transition temperature, molecular weight distribution, and percent crystallinity.) All of these important polymer characteristics have been measured and benchmarked for each PP product contact component and can be used as a quality control measure to ensure that all incoming raw materials have similar properties. This can help to ensure consistent, batch-to-batch leachables profiles for the OPUS® product contact components. The additives package has also been determined for each polymer in an effort to predict and control leachables. The platinum cured silicone components are very durable materials that are often used in medical applications. These silicone elastomers are amorphous polymers that usually do not require formulation with additives such as phthalates, antioxidants, or heat and light stabilizers. This less complex formulation greatly reduces the risk of leachables. The silicone polymers have also been fortified with silica fillers to provide the essential mechanical properties required for the OPUS® chromatography applications. As in the case of the PP components, the mechanical and physical properties of the platinum cured silicone gaskets and tubing have all been measured and benchmarked. All polymers used to construct the product contact components have met the requirements of a USP Class VI polymer and are regulated for food contact application according to 21CFR Part 177 Indirect Food Additives. The specific polymer and regulatory information is summarized in the table below.

45


Table 5.2 Polymer List and Regulatory Information

Component Material USP CFR 21 177 Animal Origin

Additives

Column Tubes 45/R – 60/R cm

70% w/w E-Glass / PP Homopolymer Composite Structure

Class VI USP <88>

177.1520 Animal Free Primary Antioxidant Processing Aid

Column Tubes 10-30 cm IDs

Extruded PP Homopolymer

Class VI USP <88>

177.1520 Animal Free Primary Antioxidant

Secondary Antioxidant

Processing Aid

Acid and Metal Scavengers

Flow Distributors 5, 10 – 60/R

Compression Molded PP Homopolymer

Class VI USP <88>



Processing Aid


Inlet and Outlet Ports, OPUS® R Plug, OPUS® R

Inside Port

Compression Molded PP Homopolymer

Class VI USP<88>



Processing Aid

Acid / Metal Scavengers

Bed Support Screens 5, 10-

60/R

PP Woven Mesh Class VI USP<88>

177.1520 EMEA 410/01

Primary Antioxidant


Hindered Amine

Processing Aid


Flow Distributor O-Rings, OPUS® R

Plug O-Ring, OPUS® R

Inner/Outer Gaskets

Platinum Cured Silicone

Class VI 177.2600 Animal Free Silica Filler

Return Line, 10-60/R

Platinum Cured Silicone PET Braiding

Class VI 177.2600 Animal Free Silica Filler

46


Testing The polymers utilized in medical and pharmaceutical applications should be compliant with the appropriate USP guidelines and it is recommended that they meet the USP Class VI testing requirements. The appropriate extractables and leachables (E&L) testing programs should be implemented for the bioprocessing materials that come into direct contact with components of the drug formulation. Three industry groups have published “Best Practices” for conducting this testing. Several presentations and publications were prepared by the Product Quality Research Institute (PQRI) for the evaluation and safety assessment of extractables and leachables in packaging for various drug dosage forms of high risk. These recommendations apply to the primary and secondary packaging associated with these pharmaceutical products. The BioProcess Systems Alliance (BPSA) and BioPhorum Operation Group (BPOG) have also published technical guideline for evaluating the risk associated with extractables and leachables specifically for single-use processing equipment.

The Repligen approach to E&L testing is that it is the responsibility of the product technology vendor to provide a technical extractable package. Leachables, on the other hand, are considered to be process specific and the responsibility of the end user to define with specific process solutions. With that in mind, Repligen has designed an extractables program to produce a robust data package by conducting extractables testing using three solvents under exaggerated conditions of contact. The materials of contact in the OPUS® Column platform:

Extruded PP (5, and 10 – 30 cm tubes only)

Polypropylene / E-Glass Composite (OPUS® 45/R – 60/R cm tubes only)

Machined Compression Molded PP

Polypropylene Mesh

Platinum Cured Silicone O-Rings

Platinum Cured Silicone Braided Tubing

Each were extracted with USP Water, 20% Ethanol, and 2.2% Benzyl Alcohol for 72 hours at 40oC. The extracts were analyzed for semi-volatiles by GC/MS, non-volatiles by LC/PDA/MS, and inorganics by ICP-MS. In addition, the neat OPUS® components were analyzed for volatiles by Headspace GC/MS. The data generated from this comprehensive series of extractables testing is detailed below.

5.4.1 Extruded PP: OPUS® 10 – 30 cm Columns Only

Volatiles by Headspace GC/MS

Extractables µg/g

Hydrocarbon <C9 1.66


Source of extractables - The hydrocarbons can be traced back to low molecular weight oligomers present in the PP from incomplete polymerization or polymer degradation reactions. The percentages of low molecular weight oligomers have been benchmarked and are monitored for the OPUS® Column polymer components to ensure a consistent extractables profile.

Semi-Volatiles by GC/MS

Extractable Water (µg/g) 2.2% Benzyl Alcohol

(µg/g)

20% Ethanol

(µg/g)

------- < 1 < 1 < 1

47


Non-Volatiles by UPLC/PDA/MS


(µg/g)

20% Ethanol

(µg/g)

Unknown 1.84 1.83 3.83

Unknown 2.86 4.15 10.19

Inorganics by ICP-MS


(µg/g)

20% Ethanol

(µg/g)

Phosphorous < 4.514 < 24.784 < 4.636

Sodium < 1.128 1.922 < 1.159

Calcium 1.858 2.780 2.086

Silicon 4.022 1.418 1.011

Source of extractables - Sodium and Calcium can be traced back to the metal stearates and oxides used to scavenge residual catalysts and acid byproducts in the polymer. Silica is due to the siloxane based lubricant used in the polymer manufacturing.

5.4.2 Machined Compression Molded PP: OPUS® 10 – 60/R cm Columns


Extractables µg/g

------- < 1



(µg/g)

20% Ethanol

(µg/g)

------- < 1 < 1 < 1



(µg/g)

20% Ethanol

(µg/g)

Unknown 2.74 < 1 1.93

Unknown 1.84 < 1 < 1

Unknown 2.75 4.14 9.86

Unknown < 1 < 1 4.38



(µg/g)

20% Ethanol

(µg/g)

Phosphorous < 4.626 < 4.766 < 23.588

Sodium < 1.157 < 1.191 1.414

Calcium 2.233 1.630 2.197

Silicon 3.615 1.687 < 1

48



5.4.3 PP Mesh: OPUS® 10 – 60/R cm Columns


Extractables µg/g

------- < 1



(µg/g)

20% Ethanol

(µg/g)

------- < 1 < 1 < 1



(µg/g)

20% Ethanol

(µg/g)

Various Pegylated

Compounds

1 – 150 1 – 108 1 - 315

Unknown < 1 1.12 2.06

Unknown 1.50 2.62 2.42

Unknown < 1 < 1 3.84

The Pegylated extractables displayed a characteristic mass distribution of 44 amu increments. The empirical formulae for the prominent ions observed for these compounds ranged from C15H33O8 - C40H82NO13.



(µg/g)

20% Ethanol

(µg/g)

Phosphorous < 4.797 < 24.385 < 4.95

Sodium 2.739 5.090 2.075

Calcium 1.855 2.322 1.829

Silicon 3.233 1.474 < 1


49


5.4.4 P/ E-Glass Composite: OPUS® 45/R – 60/R cm Columns Only


Extractables µg/g


Source of extractables – The hydrocarbons can be traced back to low molecular weight oligomers present in the PP from incomplete polymerization or polymer degradation reactions. The percentages of low molecular weight oligomers have been benchmarked and are monitored for the OPUS® column polymer components to ensure a consistent extractables profile.



(µg/g)

20% Ethanol

(µg/g)

------- < 1 < 1 < 1



(µg/g)

20% Ethanol

(µg/g)

Pegylated

Compound

< 1 < 1 1.13

Unknown 24.15 < 1 < 1

Unknown < 1 < 1 1.01

The Pegylated extractables displayed a characteristic mass distribution of 44 amu increments. Two mass distributions within the single chromatographic peak were detected. The empirical formulae for the prominent ions observed for these compounds ranged from C15H33O8 - C19H44NO10.

The unknown peak in the water extract was detected by LC/PDA. UPLC with fraction collection was used to isolate this unknown. The fraction was evaporated to dryness and re-analyzed by both GC/MS and LC/MS however no observable MS spectrum could be observed. NMR analysis of the fraction was inconclusive since significant signal could not be generated.


Extractable Water (µg/g) 2.2% Benzyl Alcohol (µg/g)

20% Ethanol (µg/g)

Boron 1.110 1.941 1.323

Phosphorous < 23.128 28.175 27.841

Sodium 6.624 5.822 4.413

Calcium 8.488 9.445 9.415

Silicon 25.994 30.921 21.182

Source of extractables – Sodium and Calcium can be traced back to the metal stearates and oxides used to scavenge residual catalysts and acid byproducts in the polymer. Silica, Boron, and Phosphorous are common elements found in borosilicate and borophosphosilicate glass.

50


5.4.5 Platinum Cured Silicone O-Ring: OPUS® 10 – 60/R cm Columns


Extractables µg/g

------- < 1



(µg/g)

20% Ethanol

(µg/g)

------- < 1 < 1 < 1



(µg/g)

20% Ethanol

(µg/g)

Unknown < 1 < 1 3.00

Unknown 1.33 1.87 4.32



(µg/g)

20% Ethanol

(µg/g)

Potassium < 0.454 < 0.442 9.989

Phosphorous < 4.710 < 22.482 < 4.858

Sodium < 2.272 1.579 < 1.073

Calcium 1.183 1.370 1.075

Silicon 19.344 15.626 19.365

Source of extractables – A silica based filler is used to provide enhanced mechanical properties to the platinum cured silicone polymer. The filler is the primary source of inorganic based extractables from this component.

5.4.6 Platinum Cured Silicone Braided Tubing: OPUS® 10 – 60/R cm Columns


Extractables µg/g

Decamethylcyclopentasiloxane 1.09

Source of extractables - Decamethylcyclopentasiloxane is most likely present in the low molecular weight silicone based processing lubricant used during tubing manufacturing.

51




(µg/g)

20% Ethanol

(µg/g)

------- < 1 < 1 < 1



(µg/g)

20% Ethanol

(µg/g)

Unknown 30.73 < 1 < 1

Unknown < 1 < 1 3.86

Unknown < 1 < 1 1.91

Unknown 1.52 2.66 6.95

The unknown peak in the water extract was detected by LC/PDA. UPLC with fraction collection was used to isolate this unknown. The fraction was evaporated to dryness and re-analyzed by both GC/MS and LC/MS however no observable MS spectrum could be observed. NMR analysis of the fraction was inconclusive since significant signal could not be generated.



(µg/g)

20% Ethanol

(µg/g)

Phosphorous < 4.710 < 22.482 < 4.858

Sodium 51.538 4.316 5.344

Calcium 1.269 1.316 1.561

Silicon 26.584 36.007 53.586

Source of extractables – A silica based filler is used to provide enhanced mechanical properties to the platinum cured silicone polymer. The filler is the primary source of inorganic based extractables from this component.

5.4.7 Conclusions

The product contact materials used in OPUS® pre-packed chromatography columns meet the requirements of a USP Class VI plastic and are regulated for food contact applications. Each polymer has been hand selected based on its mechanical / physical properties, inherent stability, and low potential risk for leachables. Only those polymer additives that are functionally necessary are present in the OPUS® product contact components. This has been confirmed experimentally through a complete quantitative analysis of the additives package in each polymer. Extractables testing has been conducted for the product contact components using conditions of 40ºC for 72 hours in three solvents: USP water, 20% ethanol, and 2.2% benzyl alcohol. The extracts were analyzed for volatiles, semi-volatiles, non-volatiles, and metals. The identified extractables were correlated back to their source of origin in the OPUS® component so that they can be controlled during manufacturing. Structure elucidation was performed on the unknowns and empirical formulae were proposed when possible. The physical properties of the polymers that have the most influence over leachables (such as melting point, glass transition temperature, percent crystallinity, and molecular weight distribution) have been benchmarked to ensure a consistent batch-to-batch leachables profile for these components and a well-managed supply chain for raw

52


materials used in their construction. The result is a chromatography column which has achieved a minimization and control of leachables through comprehensive material characterization, quality control, and extractables testing.

5.5 Leachables: OPUS® Column Strategy The extractables information in this regulatory support file is designed to be the foundation of a leachable substances therapeutic safety assessment. The data presented represent an extractables profile of what might be extracted from an OPUS® Column under extreme exaggerated conditions (prolonged exposure at high temperature).

Role of the End User Each process is different, and therefore must be evaluated for impact of leachables. In many cases, the data provided by an extractables test can be used to create a risk based assessment to support a leachables program.

Role of Repligen Repligen will provide suitable component test materials and upon request to support application and process specific leachables testing.

5.6 Considerations for Leachables Testing Purpose To assess the safety risk posed to patients through exposure to low levels of chemical entities extracted from plastics in the manufacturing process.

Role of the “model solvent” extraction test In general, the model extraction vehicles in a standard extractables test will likely show all of the materials which could possibly be extracted from a plastic product. Model stream extraction vehicles are chosen for their ability to extract compounds from plastics as well as their relevance to the test application (chromatography in this case). The results of extractable tests can be compiled and analyzed to create a risk based approach for conducting (or not conducting) a leachables study.

Chromatography: An assessment of application impact In the case of chromatography the method from start to finish should be considered, however, a risk analysis should focus on the elution fluid, which in most cases will be an aqueous solution. Although a pre-packed chromatography column will see solvents such as 0.5N NaOH, phosphoric acid, high and low pH additives, and high salt conditions, in general none of these compounds or additives are meant to pass into the final product pool. In addition, impact to the final product pool is limited due to multiple chromatography or filtration steps which help to eliminate extractables throughout a typical downstream process. Therefore, to assess the impact of other solvents in addition to the elution solvent, extractables and leachables studies can be carried out with the proposed purification methods in mind. This will enable a thorough understanding of the impact of the chromatography steps as it pertains to leachables in the final product pool.

53


5.7 References

“Safety Thresholds and Best Practices for Extractables and Leachables in Orally Inhaled and Nasal

Drug Products”, Product Quality Research Institute, Arlington VA, September 8, 2006

D. Jenke PQRI PODP Workshop Feb.22-23, 2011

PQRI Review Article – PDA J Pharm Sci and Tech 2013, 67 430-447

Bio-Process Systems Alliance: www.bpsalliance.org . Formed in 2005 as an industry-led corporate

member trade association dedicated to encouraging and accelerating the adoption of single use

manufacturing technologies used in the production of biopharmaceuticals and vaccines.

Parenteral Drug Association: www.pda.org . The leading global provider of science, technology and

regulatory information and education for the pharmaceutical and biopharmaceutical community.

Founded in 1946 as a nonprofit organization, PDA is committed to developing scientifically sound,

practical technical information and resources to advance science and regulation through the

expertise of its more than 9,500 members worldwide.

Recommendations for Extractables and Leachables Testing - Part 1: Introduction, Regulatory Issues

and Risk Assessment

Bioprocess Systems Alliance; Bioprocess International 5(11): p36-49 December 2007

Recommendations for Extractables and Leachables Testing - Part 2: Executing a Program

Bioprocess Systems Alliance; Bioprocess International 6(1): p44-53 January 2008

USP General Chapters <88> Biological Reactivity tests, In Vivo

Toxicon USP <88> “Class Tests

Estimation of toxic hazard – A decision tree approach. G.M. Cramer and R.A Ford

Fd Cosmet.Toxicol. Vol16 pp.255-276 Pergamon Press 1978

Threshold of Toxicological Concern Concept in Risk Assessment. R Kroes, J Kleiner and A Renwick

Toxicological Sciences 86(2), 226-230; 2005

Parenteral Drug Association: Assessment of E/L from Bags, Tubings and Filters

http://www.pda.org/

54


6. Appendix

6.1 OPUS® 5, 10-60/R cm ID Material Certificates (available upon request)

Refer to Section 2 for a summary of the product contact materials. The following documents are

available upon request. Please email our Customer Service department at [email protected] for

more information:

5, 10-30 cm ID Column Tube, 10-60/R cm ID Flow Distributors, Inlet Port, Return Port, and Outlet

Port: Class VI Documentation

5, 10–30 cm ID Column Tube, 10–60 cm ID Flow Distributors, Inlet Port, Return Port, and Outlet

Port: Animal Origin Documentation

5, 10–30 cm ID Column Tube, 10–60 cm ID Flow Distributors, Inlet Port, Return Port, and Outlet

Port: Component Quality Documentation (21 CFR 177)

5, 10–60 cm ID Bed Support Screens: Class VI Documentation

5, 10–60 cm ID Bed Support Screens: Animal Origin Documentation

5, 10–60 cm ID Bed Support Screens: Component Quality Statements (21 CFR 177)

5, 10–60 cm ID O-Rings: Class VI Documentation

5, 10–60 cm ID O-Rings: Animal Origin Documentation

5, 10–60 cm ID O-Rings: Component Quality Documentation (21 CFR 177)

10–60 cm ID Return Line: Class VI Documentation, Component Quality Statements (21 CFR 177)

10–60 cm ID Return Line: Animal Origin Statements

45-60 cm ID Tube: Animal Free Statement

55


6.2 Engineering Drawings

6.2.1 OPUS® 5 cm Column

56



57


6.2.3 OPUS® 14 cm ID Column

58



59



60



61


6.2.7 OPUS® 45 cm and 45R Column

62


6.2.8 OPUS® 60 cm and 60R Column

63


6.3 Packaging

6.3.1 OPUS® 5 cm Column Packaging

Box design

Packaging inserts

64


6.3.2 OPUS® 10 - 14 cm Column Packaging

65


6.3.3 OPUS® 20 - 30 cm Column Packaging

66



67



68


Index

B

Bioburden.......... 3, 5, 10, 25, 27, 31, 35, 36, 37

C

Certificate of Analysis .. 3, 5, 10, 11, 12, 13, 16, 36, 37

Cleanability.................................................... 26

E

Endotoxin .......... 3, 5, 10, 25, 27, 31, 35, 36, 37 Extractables .... 3, 44, 45, 47, 49, 50, 51, 52, 53,

54, 56

G

GMP Run Ready . 3, 5, 8, 10, 12, 13, 15, 34, 35, 36, 37

L

Leachables ............... 3, 4, 44, 45, 47, 49, 55, 56

M

Material Safety Data Sheet ....................... 3, 29 Materials of Construction ... 3, 5, 19, 20, 21, 22

P

Packaging .................... 4, 16, 66, 67, 68, 69, 70 Pre-GMP ................................ 3, 5, 8, 10, 11, 37

Q

Quality ........... 3, 7, 8, 16, 29, 30, 33, 49, 56, 57

S

Shipping .............................. 3, 5, 10, 16, 38, 39 Stability ......................... 3, 5, 19, 24, 25, 26, 43

W

Warranty ................................................... 3, 27 Work Order .... 5, 11, 12, 14, 15, 16, 28, 32, 33,

34, 35, 36