di bio gmp education itb apr 01 2014

5/21/2018 DI Bio GMP Education ITB Apr 01 2014

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Introduction of Biotechnologyand Manufacturing

Suh, Chang Woo PhD.April 01, 2014

PT. Daewoong Infion


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Lecturers

- 1998.03 ~ 2002.02 : PhD in Biochemical Engineering, University of Hanyang

- 2002.03 ~ 2004.02 : Post Doc. in Microbiologics, Microbiochip Center, Korea

- 2004.02 ~ 2012.6 : Daewoong Bioresearch center for development of

EGF, EPO, hGH, BMP-2, and BTA

- 2012.07 ~ Present : Global business development for biologics.

Establish new biological GMP in Indonesia.

Name: Suh, Chang Woo

Department: PT. Daewoong Infion

Telephone : +62-813-3324-2224

E-mail: [email protected]


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Biotechnology & Future

1) What is biological products?

2) History and future of Biotechnology

How to make Biological Products2) Manufacturing process

3) Quality Control of Biologics

Introduction of PT. Daewoong Infion

1) Mission / Goal / History

2) Product Information of EPOSIS

3) Progress of construction

CONTENTS


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PART 1.

BIOTECHNOLOGY & FUTUREWHAT IS BIOLOGICAL PRODUCTS?HISTORY AND FUTURE OF BIOTECH.


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Biologicalis a medicinal product such as a vaccine, blood or blood

component, allergenic, somatic cell, gene therapy, tissue, recombinant

therapeutic protein or living cells that are used as therapeutics to treat diseases.

Cultured cartilage

Heparin

Growth Hormone

Albumin

rh-EPO

Human blood

Red blood cell

Cord blood

Kidney

Bone

Heart,

cornea

Steam Cell

Cultured organ

Vaccinie

mAb

Tissue

Gene therapy

Definition of biological Products


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Modern Biotechnology from DNA

James Watson & Francis

Crick (1953)

Englishmen responsible

for the discovery of thedouble helix structure of

DNA using X-ray

photographs


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Wow!! Recombinant Technology

Paul Berg (1972)

Stanford University scientist who first developed recombinant DNAtechnology, a method for insertion of genetic material from one

organism into another.


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Stem Cell!! Magic but Dangerous

Stem cellsare undifferentiated biological cells that can differentiate into

specialized cells


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Clones will come to our world!

Ian Wilmut (1996) Dolly is the first animal cloned from diploid cells is

produced in Scotland


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Are you origin or copy?

Dolly gave birth to four

lambs

Dolly the sheep, the first mammal to be cloned from an

adult cell
http://www.time.com/time/magazine/0,9263,7601970310,00.html


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How do you think about Human Clone?

Bush says cloning human embryos is 'morally

wrong.'

Movie: The Island (2005)


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Are you agree with it?

Photo of mouse growing a "human ear"

- a shape made of cartilage


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Human can join with Machine.

knowledge from the joints, muscles, and nerve


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PART 2.

BIOLOGICAL PRODUCTS

FIVE FEATURES FOR A BIOTECH. DRUG

MANUFACTURING PROCESS

QUALITY CONTROL OF BIOLOGICS


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What is recombinant protein?

Recombinant DNA technology,joining together of DNA molecules

from two different species that are inserted into a host organism toproduce new genetic combinations that are of value

to science, medicine, agriculture, and industry.


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API Original Name Company in Korea

Nepidermin Easyef DW

Erythopoietin Eprex DA, CJ, LG, DW

Interferon alpa Intron A DA, CJ, GC, LG

Interferon gamma Immukine LG

Somatropin Genotropin DA, LG, DW

Filgrastim Neupogen DA, GC

BMP-2 Infuse DW

Neurotoxin Botox MT, HG, DW

Abciximab ReoPro Isu

Infliximab Remicade Celltrion

Korean Recombinant Protein Drugs


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Choice of Production Cell Line


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Manufacturing Process of r-Protein

Upst

ream

proce

ssing

Downstream

processing

Purif icat ion/Concentrat ion

(Ultraf i l trat ion/ion exch ange or

precipitat ion)

Sealing/Labell ing

/pac kag in g

Generation of Cell Substrate

Preparation of ExpressionConstruct

Cell Banking

Cell Culture/Fermentation

Purification/Isolation

Drug Substance

Formulation

Drug Product

Propagation of WCB

Starter cultu re

Production-scale

culture/bioreactor

Recovery

(Centr ifug ation or fi l trat ion)

Chromatographic p ur i f ication

Addi t ion of excip ients

Steri le fi l trat ion/

Asept ic f i l l ing

Adjustment potency


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Key Point of Biological Products

Expression system: new cell lines Mamalina cell lines: regulation of syclin-dependent kinase, anti-apoptic

proteinBCl2, PRC-6

Yeast strain humanized for N-linked glycosylation

Other alternative expression systems:

Insect cells (SF9, Hi5) Transgenic plants (Maize, tobacco, potatoes)

Transgenic animals (goat, sheep, rabbit, )

Production system: Serum free media Cell-culture systems

Structure and post-processing modifications Impact on yield and consistency of production


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DNAplasmid for hEGF

Recombinant E.coli

for production of hEGF

E.coliE.coli

EGF

EGFEGF

EGF

Recombinant hEGF

Cloning

Fermentation

DNA synthesis

by chemical reaction

E.coli

10 NH3 2CH4

4C2H6

2 H2O

Production Process for rhEGF


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Cell Culture

Fluid

Fermentation

Cell Removal by microfiltrationFiltrationChromatographyFiltration

Chromatograpahy Chromatograpahy Filtration Lyophilization rhEGF

E.coli


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Protein Purification

1. Molecular weight Ultracentrifugation

Dialysis

Gel filtration

SDS PAGE

2. Charge Ion Exchange Chromatography

Native gel electrophoresis

Isoelectric focusing

3. Affinity

Exploited with cloning

His-tagged proteins purified on Nickel columns.

GST fusion proteins purified on glutathione columns.

4. Solubility


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SDS-PAGE

1. Catalase, cytochrome C,

a-lactalbumin

2. Hemoglobin, Cytochrome

C, a-lactalbumin

3. BSA, cytochrome C,a-lactalbumin

4. Hemoglobin, myoglobin,

a-lactalbumin

5. Ferritin, cytochrome C,

a-lactalbumin6. Ferritin, myoglobin,

a-lactalbumin

1 2 43 5 6

lighter


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Gel Filtration (SEC)

The largest molecules are eluted from the column first and the smallest are elutedlast. Molecular weight range is dependent upon the pore size of gel matrix


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Ion Exchange Chromatography

UVabsorbance Increasing salt

concentration


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Affinity

Immunoaffinity chromatography - antibody

Metal affinity chromatography - chelation

Dye affinity chromatography - dye tendency

The most expensive, the mostvarious, the most excellent

The affinity between a ligand

with a high specificity and the

protein of interests. (or a

particular impurity)

Use early in a purification

scheme

Ex, Protein A affinity :

Mab - capture stage


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Quality Control for Biological Products

Genetic Stability

Cell Substrate

Viral Safety

Preclinical Safety

Generation of Cell Substrate

Preparation of ExpressionConstruct

Cell Banking

Cell Culture/Fermentation

Purification/Isolation

Drug Substance

Formulation

Drug ProductProduct Stability

Specifications


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Methodology of Characterization

Safety LAL test, rabbit pyrogen test, bacterial culture methods

Purity & Characterization including but not limited to: Reversed-phase HPLC, Peptide mapping, MS

SDS-PAGE, Western analysis, capillary electrophoresis

SEC, AUC, FFF, light scattering

Ion Exchange Chromatography

Carbohydrate analysis (capillary electrophoresis, HPAEC = high-pH anion-exchange chromatography, IEF for sialic acid)

Identity N-terminal sequencing

Peptide mapping

Immunoassays (ELISA, Western blotting)

Potency Animal-based assays, cell-based assays, reporter gene, biochemical (e.g.,

enzyme activity)

Protein content RIA, ELISA, UV absorbance, Bradford


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1. Expression System

Identify, isolate and clone the gene coding for thedesired protein

Construct a vector containing: The gene

The expression controls (promoter, secretion signal)

Insert the vector into the selected micro-organism Escher ichia col i

Saccharomyces cerevis iae

Mammalian cells

Genetically modified plants


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2. The Production System

PurposeOptimize survival conditions for the genetically modif

ied microorganism

So that it produces the desired protein

With acceptable yield

Materials & Methods

Selection of cell culture medium

Selection of culture conditions

Selection of culture equipments:fermentor, cytocultor


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3. The Purification System

Purpose extract protein from a complex growth medium

Achieve close to 100% purity

Without altering the protein

Materials & Methods

Sequence of purification steps

Filtration/ultrafiltration

Precipitation/resolubilization Chromatography (ion-exchange, affinity, etc.)


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4. Nature of Active Product

Proteins Chains of amino-acids

Sequence of amino-acids encoded by genes

Folded into 3-D conformation

To obtain biological activity Host-dependent post-translational modifications (su

gars)


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5. Pharmaceutical Formulation

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Purpose

Maintain biological activity

By maintaining active protein conformation in sol

ution

Materials & Methods

Stabilization (albumin, glycerol)

Storage at low temperature


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Part 3.

Introduction of PT. Daewoong Infion

Mission / Goal / History

Product Information of EPOSIS

Progress of construction(Utility, AHU, Panel, Paint, Pipe, Electric)


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Mission of PT. Daewoong Infion

1. Contribute to the development of the Indonesian Biologics Technical transfer of Biologics to Indonesian Pharmaceutical company

Co-development with governments and hospitals of Biosimilar and new biologics

2. Contribute to the health of the Indonesian people.

Medical technology & skill transfer through Doctor to Doctor and Doctor training

education

Medical industry Development

3. Respected company to all Employees, Partners, Customers

Everyone WinWin

Provide the right information & optimized solution to the customers

Priority to the growth of employees than the development of the company


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Goal of Manufacturing Biologics

To be the first and the best Bio GMP factory inIndonesia

Provide more affordable biological products for theIndonesian people

Provide training program development,standardization and bio-GMP expert educationprogram for contributing to the biotechnology industryin Indonesia

Prototype of Biologicals manufacturing in Indonesia

Contribute to Indonesian government income (devisa)


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History of PT. Daewoong Infion

1. Progress:

April 2012: Contract of JVC. July 2012: Approval of investment from BKPM.

Feb. 2013: Approval of DOE, Domicile and NPWP.

May. 2013: Approval of GMP Drawing from BPOM.

2. Concept & Basic Design

Total area (1st+2nd): 2,550 m2

API (630 m2), Product (620 m2), QC (230 m2), non-control area (1,070 m2).

3. Initial Total Investment : USD 20 MIO


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POS S Recombinant Human Erythropoietin Pre-filled Syringe Inj.


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Can we see EPO?

EPO is a Protein which is a hormone in human body.

Other names include:

Erythropoetin

Epoetin

Hematopoietin

Hemopoietin

Erythropoietin Signal and Action


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Mechanism of action

The feed-back regulation of EPO synthesis

Ref.) Holes human anatomy and

physiology 10thedition

EPO regulates the

differentiation and

proliferation

of erythroblast

precursor cells


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Indications & Complications

Indications Acute & chronic renal failure

Drug poisoning

Refractory edema

Metabolic Disturbances Complications

Low blood pressure

Fatigue

Nausea Iron-deficiency Anemia : blood loss from the tech

nique of hemodialysis


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Where is my blood?

Chronic KidneyDisease

CRF, Cancer, Dialysis, HIV,Infection and other cause

Anemia

TransfusionEPO

Treatment

Infection

est Choice


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Progress of Construction

(a) Frame for Panel

(c) Floor & Painting

(b) Ducting & Piping

(d) Mushola


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Progress of Construction

(e) pure water generator

(g) electric generator

(f) WFI generator

(h) chiller


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Education of Human Resource

Director of Factory

Manuf. QA QC Eng. Admin.

Head

Culture

Purification

Formulation

Filling

Packaging

Head

Lot release

Validation

AQR

SMF/SOP

RegulationDocument

CAPA/OOS

Changing

Process

Education

Head

Chemical test

Biological test

Raw materials

Microbial

Sterile testAnimal test

Stability

Equipment

Head

Machinery

Equipment

AHU/HVAC

Clean room

Electric/pipeWater/WFI

Head

Ware house

Production

Accounting

HR manager

BD/Export

GMP Committee


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Tech. Transfer of Erythropoietin

1. Technical Transfer of Manufacturing

1) Establish of Cell Line in Korea WCB (working cell bank) from RCB (research cell bank)

Qualification of WCB

2) Media Fill in Daewoong Infion

Conducting Media Fill Test before PV production

3) PV production in Daewoong Infion

PV 3 batch Long term stability test on going for two years


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Tech. Transfer of Erythropoietin

2. Technical Transfer of Analytical Method SOP training

Calibration Training for Equipment

Method Validation

Animal Management and Testing Training


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Formulation research

Global First/Advanced generics

R&D out-sourcing

Hyderabad, India

Product development

for domestic market

Botanical Medicine

Beijing, China

Yongin, Korea

150 Scientists

Well-balanced research areas;- NCE/Biologics

- Generics/Incrementally modified drugs

- Botanical Medicines

Active scouting of external innovation

DaewoongR&D Centers


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Pharmtech Dec. 2002

Annual salaries as job function


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Terima Kasih !

Always Smile with Daewoong Infion

di bio gmp education itb apr 01 2014

Documents

new biological gmp

daewoong infiontelephone

human ear

human clone

daewoong bioresearch

diploid cells

cloning human embryos

modern biotechnology