usp standards for biologics tina s. morris, ph.d., vice president, biologics & biotechnology...
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USP Standards for BiologicsTina S. Morris, Ph.D., Vice President, Biologics & Biotechnology USP-NF
Fouad Atouf, Ph.D., Director, Biologics & Biotechnology, USP-NF
User ForumJanuary 17th, 2013Istanbul, Turkey
USP Standards—Biologicals
Major B&B Themes and Initiatives for 2010-2015
Modernization of MonographsOutreach to Industry for New MonographsProcedural Chapters and Characterization/Measurement of
Ancillary and Process MaterialsGrowing the portfolio to support all biologicsModernization of analytical approaches
Biological Potency Tests:Implementation of new USP ChaptersAssay transitions: animal to cell, cell to binding, activity to amount of
substance – questions of equivalence and units
Product Class Chapters:Monoclonal antibodiesGlycoconjugate vaccines
Vertical Standards – Monographs
Role and Use: Clearly define identity, strength and purity, as well as other
important quality attributes at the product level Allow independent testing and verification based on a public
standard
Considerations for Standard Development: Complexity of specifications and system suitability criteria Biological potency assignments and unit maintenance
– Across manufacturers– Internationally
Product- specific vs. common product class requirements For well-characterized and legacy products: inclusion and
bridging to new analytical technology
Official USP Biologics Monographs by Product Class
Product Class Number of monographs
peptide 47
enzyme 12
complex extract 11
carbohydrate 11
glycosaminoglycan 9
other 5
Tissue product 6
IgG/serum 9
Blood component/protein 5
Vaccine 3
39%
10%9%
9%
8%
4%
5%
8%
4%3%
1%
B&B Overall Monograph Distribution by Product Class
peptide enzyme complex extract carbohydrate
glycosaminoglycan other Tissue product IgG/serum
Blood component/protein Vaccine Other
Current Biologics in the US Market
From Kozlowski et al., NEJM 265;5, 2011
Biologics with no official USP Monograph
Betaseron
Novoseven
Epogen/Procrit
Lucentis
Rebif
Cerezyme
Herceptin
Erbitux
Kogenate FS
Humira
Aranesp
Rituxan
Neulasta
PEGASYS
Tysabri
Synagis
Remicade
Recombinate
Avastin
Avonex
Enbrel
2008
2010
2013
2013
2013
2013
2014
2014
2014
2014
2014
2015
2015
2015
2015
2015
2018
2019
2019
2026
2028
Patent Expiry Horizon
Current Key New Monograph Development Projects
BB1:– Insulin Glargine– Fondaparinux– Dalteparin Sodium– Epoetin– Octreotide
Soon in ballot:– Filgrastim
BB2:– Factor IX, plasma
derived
Soon in ballot:• Sipuleucel T
Current Key Monograph Modernization Projects
BB1:– Heparin Sodium, stage 3– Menotropins– Pancreatin/pancrelipase– Insulins
Horizontal Standards - Procedural
Benefits: Access to validated procedures and procedure performance
criteria early in development
Solid anchor point for product characterization
Facilitate comparability during all development stages
Directly apply ICH guidance, including:– Q5E– Q6B
Establish consistency in analytical expectations
From General to Specific - Procedures
<1084>Glycoprotein and
Glycan Analysis – Introduction and Choice of Analysis Methods
<212>Oligosaccharide Analysis
<210>Monosaccharide Analysis
Monosaccharide Mixes 1–4
Oligosaccharide mixturesfrom - Human α acid
Glycoprotein- Fetuin- RNAse B- Human IgG
Single Monosaccharides
Guidance & Information
Procedures & Performance
Criteria
Procedural (Horizontal)
Standards for System Suitability
& Validation
<1084> - Glycoprotein Analysis Strategies
<1084> Glycoprotein Analysis Strategies
Test Chapters – Current Major Initiatives
Impurities
• <30> Residual DNA
Physicochemical Tests• <212> Oligosaccharide Analysis
• <210> Monosaccharide Analysis
• <121.1> Insulin Physicochemical Analysis
• <209> Low Molecular Weight Heparin Molecular Weight Determinations
• <XXX> Collagen
Potency Assays and Content
Measurement• <57> Protein Determination Procedures
• <123> Glucagon Bioidentity Tests
• <124> Epoetin Bioassays
• <126> Somatropin Bioassays
• <208> Anti-Factor IIa and Xa Assays for Unfractionated and Low Molecular Weight Heparins
Vaccine Chapters
<1235> Vaccines for Human Use (completed)
<1238> Bacterial Vaccines (completed)
<1239> Viral Vaccines (in progress)
<XXXX> Toxins/Toxoids
Sub-<1000> Analytical Chapters for Key Quality Attributes and RS
Possible PC Chapters
<1234> Polysaccharide and Glycoconjugate
<XXXX> Live Attenuated
<XXXX> Other Subunit
<XXXX> Live Attenuated
<XXXX> Killed Viruses
<XXXX> Subunit Vaccines
<129> Analytical Procedures for Recombinant Therapeutic Monoclonal Antibodies
Will contain a collection of validated compendial procedures with established system suitability criteria for therapeutic MAbs
Will be accompanied by USP MAb System Suitability RS
Will not contain product or class specific acceptance criteria
Will be supported by a suite of >1000 Information Chapters that discuss quality attributes, manufacturing and quality control aspects for MAbs
Benefits: Quality specifications and standards for complex ancillary and
process materials – Give access to process ingredients of consistent quality– Control process variability– Facilitate testing of process intermediates and bulks
Challenges in Standard Development: Fast evolution of process materials:
– Serum-free, proprietary custom media– Engineered 2nd and 3rd generation materials, e.g. Protein A
Defining key quality attributes for complex materials, e.g. Fetal Bovine Serum (FBS)
Horizontal Standards– Ancillary & Process Materials
Raw/Ancillary Materials
Raw materials may or may not remain in the final therapeutic product as active substances or excipients
– Ancillary materials are a subset of raw materials
Ancillary products may exert an effect on a therapeutic substance – (e.g. a cytokine may activate a population of cells), but they are not intended to
be in final formulation
Concerns related to raw materials qualification have been amplified
by recent materials supply issues: – Glycerin, Heparin, Melamine– Animal derived material (serum) used in manufacturing of some biologics
Some components are more critical than others! Risk assessment strategies are required to ensure quality – A critical material will come in contact with cells with a potential to alter either the
growth characteristics of the cells or the ability of the cell culture to meet lot release specifications.
Ancillary Materials Standards: USP Approach
<1043> Ancillary Materials for Cell-, Gene-, and Tissue-Engineered
Products
Specific AM Chapters
<90> FBS Quality Attributes<92> Cytokines and Growth Factors Quality Attributes
Reference Standards:- FBS- Interleukin-4- FGF-2- Transferrin- G-CSF
General Information
Chapter (Guidance)
Ancillary Material Requirements for
Specific AMs
Ancillary Material Reference Standards
Tier 1 – Low-Risk, Highly Qualified Materials with Intended Use as Therapeutic Drug or Biologic, Medical Device, or Implantable Material
Tier 2 – Low-Risk, Well Characterized Materials with Intended Use as AMs, Produced in Compliance with GMPs
Tier 3 – Moderate-Risk Materials Not Intended for Use as AMs (frequently produced for in vitro diagnostic use or reagent grade materials)
Tier 4 – High-Risk Materials, Materials not Produced in Compliance with cGMPs and materials not intended to be used in cell manufacturing
Risk-based Classification of Ancillary Materials
Risk-based Qualification – USP <1043>
Elements of Qualification and/or Risk ReductionActivities Tier 1 Tier 2 Tier 3 Tier 4
Master File cross reference X X X X
Certificate of analysis X X X X
lot-to-lot effect on process performance X X X X
Removal from finished product X X X X
Stability as stored and used in specific process X X X X
Confirm Certificate Analysis Test X X X
Vendor Audit X X X
Upgrade Manufacturing to GMP level X X
Develop internal specifications X X
Lot to lot comparability may be needed X X
Testing for adventitious agents may be needed X X
Traceability to country of origin, safety from animal diseases X
Adventitious agent testing for animal source-relevant viruses X
Recombinant Human Interleukin 4 -USP
Interleukin-4 Standard Requirement– Specific Activity: Not less than 0.5 x 107 USP Units /mg of total
protein• Labeled potency of RS will be based on bioactivity using TF-1 cell line
– Purity: 98% (SDS-PAGE and silver stain)– Identity: N-term protein sequencing (10 residues) and Western Blot
Associated Reference Standard – Lyophilized powder– Calibrated against International Standard (WHO)
How is the standard used?– Preparation of IL-4 for which the activity was determined by
calibration against the USP standard, will provide consistency in the manufacturing of the cell therapy product, by way of using the right amount of material.
Fetal Bovine Serum – Quality Attributes and Functionality Tests
Official uses of the USP FBS Reference Standard – Identification – Radial Immunodiffusion
– FBS Functionality Tests – Growth Promotion Curve• Five cell lines are recommended for use• Functionality tests are performed on 3 cell lines
○ Two from the recommended cell line list
○ Third is cell line relevant to the user’s application
USP General Chapter <90>
Fetal Bovine Serum (FBS)- USP
FBS Standard Requirements– Osmolality: 280-360 mOsm/Kg
– Total Protein: 30-45 mg/mL
– pH: 7.00 - 8.00
– Endotoxin: Not more than 10 units/mL
– Hemoglobin level: Not more than 30 mg/dL
– Identification: Radial Immunodiffusion (RID): species ID, IgG levels
– Functionality Assays (Growth Curve and Clonal Assay)
Associated Reference Standard (RS), under development– Liquid frozen, 10 mL– Collaborative study to include several laboratories to test:
• Identification (FBS sample positive for bovine IgG and content is < 500 mg/L)• Growth curve (doubling time in test sample is not less than 90% compared to
RS)
How the FBS Standard is Used: Growth Curve
Challenges: Cell Line, Cell Density, Cell Counting, Days in Culture
Three cell densities, determine viable cell counts on days 0,1,2,3,4, and 7. Select the
cell density that exhibit a growth curve with 3 phases: Lag, Log, Stationary; and linearover 3 time points or more
Use the selected cell density to assess thetest FBS side by side with the referencestandard FBS
Doubling time is estimated using a growth curve that is linear over three or more time points.
Acceptance Criteria:R2≥ 0.98Doubling time of test sample should be notless than 90% of doubling time of RS
Growth Curve for prostate cell line, WPE1-NB26-64
1.00E+03
1.00E+04
1.00E+05
1.00E+06
0 1 2 3 4 5 6 7Days in Culture
A
B
C
Growth Factors and Cytokines – additions to <92>. – Next revision adds FGF
Standards for enzymes as ancillary materials– Trypsin, others?
Second and third generation identification tests for complex, naturally derived materials:– Application of modern immunology and proteomics
approaches?
Future Directions for USP Ancillary Materials Standards
Thank you!