christine lattenmayer

a Novartis company

Late Phase Process Development by Applying Quality by Design

Optimising Biomanufacturing Processes, Brussels, Dec 2008Christine LattenmayerGroup-Head Cell Biology & Media DevelopmentBiopharmaceutical Operations, Sandoz Austria

C Lattenmayer, Optimising Biomanufacturing Processes, Brussels Dec 2008

Agenda

Sandoz Organization & General Novartis Concept

QbD concept – The 3 steps

Target – Definition & Redefinition

Process Development – Case Studies

Characterization


A Global Network for Biopharmaceutical Opera-tions…

Vacaville (US)Clin & commercial mfg

Basel (CH)Commercial mfg(dedicated to Simulect DS)

Kundl (A)•Tech dev•Clin & commercial mfg

Huningue (F)Clin & commercial mfg

Menges (SLO)•Tech dev•Commercial mfg

Schaftenau (A)•Tech dev•Clin mfg

Mammalian cell cultureMicrobial fermentation

Oberhaching and Holzkirchen (DE)•Dev., marketing


Cell Culture CoE Schaftenau/Austria

Development of cell culture processes for new biopharmaceuticals & biosimilars

Production of drug substance for clinical studies using fed-batch technology

In operation since 2004App. 130 people

3 USP/DSP production lines (3-13 m3)Development & analytical labsManagement, quality assurance & engineering


Obsolescence

Novel biopharmaceuticals

The Novartis Way of Thinking: Novel biologics and biosimilars are complementary

High quality Follow-on Biologics

Patent expiration

Biosimilars offer:• high quality • established

treatments• affordable costs

and thereby free up healthcare funds for new innovative drugs

Novel biopharmaceuticals offer:

• improved treatment

• new therapeutic opportunities

and thereby might replace older/less effective medications


QbD concept: The 3 Steps

Define target

Characterize result: ‘proof that it works’

Product, process, clinics

Development of processmeeting the target,"Quality by Design"

Same concept fornew biopharmaceuticals &

biosimilars


QbD concept: The 3 Steps - Same Concept for New Biopharmaceuticals & Biosimilars

New Biopharmaceutical BiosimilarProduct quality profile

PerformanceFacility-fitDefine target

Comparability to previous clinical DS Comparability to originator

Start from standard/ platform process, rational DoEExtensive product quality analysis

Development of process

2-step development (early/late phase)→ more time to gain experience with cell line/ product

1-step development → most efficient capacity useincreased challenge: rarely platform process useable, tight targets (e.g. glycosylation) to combine with high yield

Extensive set of state-of-the-art analytical methods: phys.-chem., biological characteristicsCharacterize

result Need to know structure-function relations better than originator


QbD concept: How to Define the Target?

TargetDefinition & Redefinition

Process DevelopmentCase Studies

Characterization

Define target





Target definition & Redefinition:Systematic, Scientific and Risk-Based Approach

• Target describes desired state of:Product quality: phys.-chem., biological, safety parametersProcess performance: yield, cost-of-goods (COGS), robustnessProcess fitting to commercial facility

• Quality-target is based on risk-assessment: Define critical product quality attributes based on their expected impact on safety and efficacy Redefinition of critical quality attributes

Initial target Redefined target

Continuous improvement over time


QbD Concept: Development of Process Meeting the Target

TargetDefinition & Redefinition

Process DevelopmentCase Studies

Characterization

Define target





Developing a Process Meeting the Targets

Start-run

USP media-screening

USP parameterscreening

DSP parameterscreening

USP fine-tune

DSP fine-tune/stabilities

Pilot-runs

USP media-/feed-development

DSP resin-screening

Vial-break

Definetarget

Time

Rational DoE

Interaction of parameters

Product qualitycontrol

Target, responsibilities, project plan, history,

status, milestonesFacility-fit

Focus on product quality & performanceDevelopment Plan Manufacturingoriented


Approaches:Uni-variate Designs:

• Screening selected single parameters at very broad ranges• Worst-case runs

Multi-variate Designs:• Full or partial factorial designs• Response surface

Demand-based:• Replace what cells have metabolised

Developing a Process Meeting the Targets: Rational DoE for Efficient USP Development


Developing a Process Meeting the Targets: Rational DoE for Efficient USP Development

• Investigation of main process parameters in screening and/ormultivariate designs

• Testing of harvest stabilities at different storage conditions in screening or full-factorial designs

• Investigation of media components or components groups for improving titer and triggering product quality into design specifications

• in screening designs• based on consumption rates • and if appropriate, in multivariate designs

• Investigation of feed components or components groups for improving titer and triggering product quality into design specifications


Case Studies

Single media-components may lead to tremendous increase of performanceIncreasing titer by combined medium and process optimizationAdjusting charge variants by USP developmentChanging glycosylation pattern by innovative cell culture conditionsOptimization of output of capture stepProcess-upscale for a sensitive mAB


Case study 1: Single media-components may lead to tremendous increase of performance

Early Process:commercial medium

• undefined composition• containing peptones

low product activityhigh process variationlong main-stage (3 weeks)

Final Process:in-house medium

• chemically defined• peptone-free

high product activityshort main-stage (2 weeks)

Baseline1st Improve-

ment 2ndImprove-

ment

0

2

4

6

8

10

12

14 Higher concentration

of 1 trace metal

Product-activity

[arbitrary units]

Higher concentration of 1 vitamin

> 10-fold increase in product activity

• broad media screening

• single component screening (uni-variate DoE)

Higherconcentration

of supplement 2

Higherconcentration

of supplement 1


Tite

r

Pilot PhII

ManufPhII

Osm Buffer/pH Feed

Phase II Process:FB with Peptone Feed

Phase III Process:FB with chemically defined Feed

Investigation of • Buffers / pH-regulation • Medium supplements• Feeding• Osmolalities • Seeding densities • Temperature shift • Aeration• Shaking speed / Power input

Case study 2: Increasing titer by combinedmedium and process optimization

SEC:PhIIPhIII

CEX:PhIIPhIII

Product quality on capture eluate level


Development of a biosimilar product

Define target

Confirmation: Comprehensive

comparabilityexercise

Development of biosimilar product,"Quality by Design"

Physicochemical and biological

characterization

Process development

Refinement of target,

Identificationof CQA's


Case Study 3: Adjusting charge variants by USP development

Charge-variants of mAb, analyzed by CEX, are typical product-related substances or impurities :

• acidic variants (e.g. de-amidation of Asn, cysteinylation), • basic variants (e.g. amidation of Pro, Lys-variants at C-terminus)• N-Terminal Cyclization (e.g. pyroglutamate at N-terminus)

Adjustment via DSP is partially possible, but reduces yield

Charge-variants can be adjusted via USP:• media components• process parameters


0

Varia

nt (%

)

Process parameter 2

0

Varia

nt (

%)

Process parameter 1

Case Study 3: Adjusting charge variants by USP

Analysis of charge variants using cation exchange chromatography

0K 1K 2K1Q 2Q

CPB

pE pE pE pE Q pE pE pEpE pE pE pE

K

Q pE pE QpE pE pE pE

KK

1K0K 2K1Q 2Q

Targeting charge-variants via process parameters

acidic

0

Varia

nt (%

)Process Parameter 3

Referenceproductrange




NP-LC of 2AB-labeledglycans

G0 G1 G2

Case Study 4: Changing glycosylation pattern byinnovative cell culture conditions

0,0 0,2 0,4 0,6 0,8 1,0 1,2 1,40,0

0,5

1,0

1,5

2,0

Media Component A

Med

ia C

ompo

nent

B

Reference Product Range

Media components result in desired galactosylation state in dose-dependentmanner

0

1

2

3

4

5

6

d7 d8 d9 d10 d7 d8 d9 d10

Concentration 1 Concentration 2

% M

anno

seX

stru

ctur

e

Other media-components allow targeting of fucosylation or mannosylation


Case Study 5: Optimisation of output of capture step

Estimated Response Surfacebuffer capacity _ elution buffer=50,0

3,2 3,3 3,4 3,5 3,6 3,7 3,8pH _ elution buffer

4,54,74,95,15,35,5

pH _ wash buffer

0,30,40,50,60,70,80,9

Des

irabi

lity

Desirability0,0-0,10,1-0,20,2-0,30,3-0,40,4-0,50,5-0,60,6-0,70,7-0,80,8-0,90,9-1,0

⇒ choice of conditions according to multiple response optimization:

pH of wash buffer

pH of elution buffer

buffer concentration of elution buffer

Output parameter:

HCP level in eluate

purity

step yield

elution volume

Input parameter:

pH of wash buffer

pH of elution buffer

buffer concentration of elution buffer

Improvement of washing and elution conditions of platform process conditions


Case study 6: Process-upscale for a sensitive mAB

10,0 12,5 15,0 17,5 20,0 22,5 25,03

- mai

npea

k - 1

9,65

0

pink: referenceblack: freshly thawed DS

(directly frozen after production)blue: same as shown in black, 24h open

(with air contact) at RT

Process changes• new cell line • new medium• different USP/DSP process• 7-fold higher titer

The problem: CEX analysis of DS resulted in OOS

1 cell-free harvest, fresh

2 cell-free harvest, stored

3 Capture eluate

4 Drug substance

The reason:

Non-red. SDS-PAGE:reduction of Cys-bridges during storage of capture-load

1 2 3 4

Questions:• What is root-cause for mAb-reduction?

Antibody reduction starts in PS.F, no full mAB is present in Capture E luateNo reduced antibody was observed in DS generated during process dev.

• How can mAb reduction be prevented?• Why was problem not apparent during process development?


Case study 6: Process-upscale for a sensitive mAB

Root-cause for mAB reduction:• storage of cell-free harvest under N-atmosphere• cellular enzymes, redox potential of spent medium, pH, sensitivity of mAb

How can mAb reduction be prevented?• storage of cell-free harvest at 4°C or under air-atmosphere completely prevents mAb-reduction• shorten intermediate holdtime by use of larger column for capture-step 1 PS.F 0d

2 PS.F N2, 5°C (2-3°C), 3d3 PS.F N2, RT, 3d

1 2 3

Why was problem not apparent during process development?• development was performed in air-atmosphere • lesson learnt: intermediate hold-times/conditions (temperature, air/nitrogen) are simulated at small-scale for all products

√


Developing a process meeting the target: Adjusting product quality of mABs

Main critical product quality attributes of mABs:

Biologicalactivity

Charge variantsGlycoforms Aggregation/

Degradation

USP DSP


QbD concept: Development of process meeting the target

Target Definition & Redefinition

Process DevelopmentTools & Case Studies

Characterization

Define target





Characterize process: Proof that it works

Science in combination with quality risk analyses ensure better understanding of manufacturing process and its product

Sum of process parameter ranges and their interactions ensure product quality within target specification

Mainly done at small-scale with qualified small-scale model

Process characterization = Definition of design space of final process

Design-Space (ICH Q8): = multidimensional combination and interaction of input variables and process parameters that have been shown to provide a reasonable assurance of product quality; Process changes within the design-space are not considered as ‘Regulatory Changes’ .


Validate process: Proof that it works

Process validation:

Follows process characterization

Large-scale process consistency runs at set-point

Possibly large-scale runs at extremes of design space (e.g. holdtimes, selected parameters?)

Virus clearance validation (new and used resin)

Resin re-use studies


Define target




Challenging, butefficiently deliveringthe best process and

product quality

QbD concept: The 3 steps


THANK YOU FOR YOUR ATTENTION !!!

Many thanks toMany thanks toBPO Sandoz/ Schaftenau

‚Cell Culture Center of Excellence‘Florian Unterluggauer

Heiko MeentsJörg Windisch

Special Special thanksthanks to:to:Julia Schmutzhard, Corinna Sonderegger and our CMD team

Josef Stettner and our BPD teamThomas Neumeier, Susanne Richter and our DSP team

Wolfgang Gutleben and our analytical group

christine lattenmayer

Documents

optimising

wash buffer

reference

triggering

adjusting

variate designs

biopharmaceuticals

product quality