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Quality Control in Biotechnology Quality Control in Biotechnology

Quality Control in Biotechnology - PowerPoint Presentation

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Quality Control in Biotechnology - PPT Presentation

Andrew Lees PhD Scientific Director Fina BioSolutions LLC wwwFinaBiocom Fina Chemical drugs vs Biologicals Chemical drugs can be precisely defined Physical chemical characterization ID: 461281

conjugate bulk content protein bulk conjugate protein content vaccines control amp design process size vaccine toxoid structure test product

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Slide1

Quality Control in Biotechnology

Andrew Lees, Ph.D

.

Scientific Director

Fina BioSolutions LLC

www.FinaBio.comSlide2

FinaSlide3

Chemical drugs vs

Biologicals

Chemical drugs can be precisely definedPhysical chemical characterization

NMR- structureMass Spectrometry- molecular weightChromatography- purity, quantityPotencyFormulation

Relatively easy to create “generics”Slide4

Chemical drugs vs

Biologicals

Biologicals are produced by living cellsImpossible to

control every variablecompletely characterizePrecisely replicateTraditionally,

biologicals are defined by “product by process”Product is defined by the manufacturing processQuality is compliance-driven

Goal is a “well-characterized biological”Slide5

c

urrent Good Manufacturing Process

cGMP

Doing what you said you were going to do

Proving that you did what you said

D

ocumenting that you did it.

Following SOPs

Validated methodsSlide6

Examples of Biologicals

Insulin

EPO

InterferonsToxinsAntibodyConjugates

Antibody-drug conjugatesFusion proteinsSlide7

Protein Structure

Secondary Structure:

3-dimensional

structure of

segments

Tertiary structure

:

Final

specific geometric shape that a protein assumes

Quaternary structure:

Arrangement

of multiple folded protein or coiling protein molecules in a multi-subunit complex.

Primary Structure:

Amino

acid sequenceSlide8

Insulin Slide9

AntibodySlide10

M

odifications

Post-translation modifications

Chemical changes not directly coded in DNAGlycosylation

PhosphorylationLipidationChemical degradations

OxidationsDe-amidationRearrangementsSlide11

Genetic Engineering (bacterial)Slide12

Monoclonal

AntibodiesSlide13

Bioprocessing

Unit Operations

Fermentation

Centrifugation

ChromatographySlide14

Variable inputs

Cells

“black box”

Process variables

Biologically derived

Chemically derived

Variable

c

rude product

Inherent heterogeneity

Stochastic processes

Heterogeneous

p

roduct

FormulationSlide15

Herceptin

(anti-cancer antibody)

Seven different

glycoforms

, each with different levels ofbiological

acitivity.

Variability of materialsSlide16

Quality by Design (

QbD

)

Operate within a specified design space Target Product Profile Critical Quality Attributes

Critical Product Attributes Better understanding of process and productDefining design spaceAllows for more variability

Process Analytical Technologies Real time monitoring & feedback.

Product by process

Lot release

criteria

- pass/

fail

Very difficult to make process improvementsSlide17

Design of

Experiment

Understanding the operating spaceSlide18

Design of Experiment (DOE)

A statistical method to model a process

Many fewer experiments than “one factor at a time”

Allows for modeling interactions

Useful to determine critical parameters

Critical for “Quality by Design”Slide19

Application of Design of Experiment to Conjugate

VaccinesSlide20

Design of Experiment

Concentration

CDAP Ratio

% conjugateSlide21

Process Analytical Technologies (PAT)

Real time feedback to control processSlide22

Types of Vaccines

Subunit (protein) vaccines

Tetanus toxoid

Diptheria toxoidPneumococcal (PneumoVax)

Killed vaccinesRubella, Measles, Polio (Salk)Flu

Hepatitis A

Live attenuated

vaccines

Polio (Sabin

)

Flu (

Flumist

)

Rotavirus (

Rotarix

)

Virus Like Particles (VLP

)

HPV (

Gardisil

)

New Generation Vaccines

DNA vaccines

Conjugate vaccines

Pneumococcal (

Prevnar

)

Meningicoccal

(

Menactra

)

Haemophilus

b (

Hib)Slide23

Polysaccharide Vaccine

Polysaccharide

Protein

+

Poorly immunogenic in infants

No boosting or memory

No class switching

No affinity maturation

Immunogenic in infants

Boosting and memory

Class

switching

Affinity maturation

Conjugate

VaccineSlide24

Conjugate Vaccines are EffectiveSlide25

Why Conjugate Vaccines?

Expensive

Challenging to manufacture

Many serotypes

Haemophilus

influenzae b (Hib)

Neisseria

meningiditis

Streptococcus

pneumoniae

Salmonella

typhiSlide26

HARVARD STRAIN

Cl.

tetani

INTERMEDIATE SEED

HIGB (18 -32

hrs

)

FERMENTOR

35 ± 5°C

ANAEROBIC CULTURE

Revival On ATG medium

48 hours at 37 °C

Inculcation into production

Medium (Muller &Miller)

(PH – 7.5 ± 0.1)

CRUDE TOXOID SUBLOT

STERTILE TOXIN

CULTURE HARVEST 6\7 days

SMEAR

PH

LF/ML

Sterility test

LF/ML

MLD

MTV

Antigenic purity

LF/ML

Detoxification test in mice

Detoxification with 0.5 % Formalin

4 days at Room temperature & 4 weeks at 35 °C

Millipore Filter

Clarification

Sterilization

CONCENTRATED TOXOID

BULK PURIFIED TETANUS TOXOID

POOLED CONCETRATE

(1-2 SUBLOTS)

Millipore Filter [30

KDa

] 0.2 to 0.45 micron

SUPERNATANT

STERILE FILTRATION

PURIFIED TOXOID

PRECIPITATION

Sterility test

Antigenic purity

LF/ML

Specific Toxicity

Irreversibility test

Released

for

Blending

Final Filtration (Cartridge Filter)

Dialysi

s

0.2 micron bag pore size

LF Test

LF Test

Centrifugation

(4000 rpm @45 minutes)

Precipitation with ammonium

Sulphate – I

st

salting (10% - 12%)

5

th

Day

sample collection for SMEAR, pH, growth

lysis

Precipitate with high ammonium sulphate (22% - 24%)

Centrifuge

4000 rpm

@ 1 hour

pooling

TETANUS

TOXOID PRODUCTIONSlide27

Identity

Polysaccharide composition

Moisture content

Protein impurity

Nucleic acid impurity

Pyrogen

content

Molecular size

distribution

Extent of activation

Molecular size distribution

Identity

Purity

Toxicity

Extent of

derivatisation

(if

appropriate) NR

Identity

Residual reagents

Saccharide:protein

ratio & conjugation

markers

Capping markers

Saccharide content NR

Conjugated v. free saccharide

Protein content

Molecular size distribution

Sterility

Specific toxicity of carrier (if appropriate)

Endotoxin content

Carrier Protein

Polysaccharide

Activated saccharide

Bulk Conjugate

Conjugation

Synthesis of Conjugate Vaccines

WHO Recommendations for the production and control of pneumococcal

conjugate

vaccines, ECBS, October 2003. Updated 2009.Slide28

Bulk

Conjugate1

Bulk

Conjugate2

Bulk

Conjugate3

Bulk

Conjugate4

Bulk

Conjugate5

Bulk

Conjugate6

Bulk

Conjugate7

Bulk

Conjugate8

Bulk

Conjugate9

Bulk

Conjugate10

Bulk

Conjugate11

Bulk

Conjugate12

Bulk

Conjugate13

Formulated

Vaccine

Multivalent pneumococcal conjugate vaccineSlide29

Carrier Protein

Bulk Conjugate

Final Vaccine

Polysaccharide

Activated saccharide

Identity

Polysaccharide composition

Moisture content

Protein impurity

Nucleic acid impurity

Pyrogen

content

Molecular size

distribution

Extent of activation

Molecular size distribution

Identity

Purity

Toxicity

Extent of

derivatisation

(if

appropriate) NR

Identity

Residual reagents

Saccharide:protein

ratio & conjugation

markers

Capping markers

Saccharide content NR

Conjugated v. free saccharide

Protein content

Molecular size distribution

Sterility

Specific toxicity of carrier (if appropriate)

Endotoxin content

Identity

Sterility

Saccharide content (of each)

Residual moisture

Endotoxin content

Adjuvant content (if used)

Preservataive

content (if used)

General safety test

pH

Inspection

Conjugation

Formulation

Control testing of Pn conjugates

WHO Recommendations for the production and control of pneumococcal

conjugate vaccines, ECBS, October 2003. Updated 2009.Slide30

>300 GMP steps for Prevnar13

Managing

supply

chain & supply chain quality

Each ingredient must be ready at the right time

QA/QC for bulk and formulated vaccine

Complexity of Supply Chain & Quality ControlSlide31

Thank You!

www.FINABIO.com