Microbial Services Approach to QbD and Process Characterization Case Study

Dr. Axel Erler / Lonza Ltd / 12th April 2012

World Vaccine Manufacturing Congress

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“Quality Can Be Planned.”

Joseph M. Juran

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Tox

Kno

wle

dge

Phase 1 Phase 2 Phase 3 Commercial

Process Knowledge Generation Lifecycle of a Manufacturing Process

Development Optimization Characterization Validation

Filing

Costly post-approval changes

Fixed Process

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Lonza`s New Approach to QbD andProcess Characterization in Biomanufacturing

Industry Standard

≈ 500Experimental

Runs

New Approach≈ 250 to 375

Experimental Runs

Step1

A model bioprocess with 10 unit operationsStep 2 Step 3 Step 4 Step 5 Step 6 Step 7 Step 8 Step 9 Step 10

# experimental runs for process characterization

43-70 43-70 43-70 43-70 43-70 43-70 43-70 43-70 43-70 43-70

Reduced Cost and Timeline

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Key to Keep ProcessCharacterization Cost low is ...

EfficientMultivariate

Experimental Strategies

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Process SDM

Risk Assessment

Validation

1. Main-Effect

Screening Study

2. Main-Effect

Modeling Study

Capability

Confirmatory Run(s)

Yes

No

Lonza Expands it`s DoE Toolboxby the Use of Efficient Multivariate Designs

SimulationStatistical Model

2. AugmentationNo

Yes

Effect Resolution

?~25 to 50%fewer runs

?

Process Characterization

1. Main-Effect

Modeling Study

New Approach

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Lonza`s New Approach Reducesthe Number of Required Experimental Runs

Case Study – Formaldehyde Treatment of a Purified Protein Vaccine

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Formaldehyde Treatment Reaction

Formaldehyde in Water(Paraformaldehyde)

Formaldehyde+

Protein

Crosslinked side chains(Protein aggregates)

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Process Representative Scale Down Models (SDMs)Can Be as Small as a 50mL Spinner Flask

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Lonza Continuously Expands it’s DoE Toolboxby the Use of Efficient Multivariate Designs Six input parameters only 17 experimental runs Three test levels covers non-linear effects Five center points estimates pure experimental error

1 2 3 4 5 6

Run Pattern Protein(g/L)

Quencher(Lysine-HCl, g/L)

Time (h) pH Temperature(°C)

Formaldehyde/ Protein Ratio

AggregateLevel (%)

1 ----+0 0.1 4 40 7.2 39 2.0 2.92 +-+-0- 0.2 4 56 7.2 36 1.5 14.93 000000 0.15 5 48 7.4 36 2.0 14.44 ++0-++ 0.2 6 48 7.2 39 2.5 27.55 000000 0.15 5 48 7.4 36 2.0 14.66 0+---- 0.15 6 40 7.2 33 1.5 3.67 -++0+- 0.1 6 56 7.4 39 1.5 2.58 -+-+0+ 0.1 6 40 7.6 36 2.5 8.59 000000 0.15 5 48 7.4 36 2.0 14.110 +0-++- 0.2 5 40 7.6 39 1.5 2211 -0+--+ 0.1 5 56 7.2 33 2.5 4.212 000000 0.15 5 48 7.4 36 2.0 14.213 --0+-- 0.1 4 48 7.6 33 1.5 2.714 000000 0.15 5 48 7.4 36 2.0 15.315 0-++++ 0.15 4 56 7.6 39 2.5 28.216 ++++-0 0.2 6 56 7.6 33 2.0 30.717 +--0-+ 0.2 4 40 7.4 33 2.5 31.9

Input ParametersOutput

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ProteinHCHO / ProteinTemp.pHLysineTime(Lysine-10)*(pH-7.4)(pH-7.4)*(Temp.-36)(Time-48)*(pH-7.4)

Term16.00

9.336.646.15

-5.183.22

-2.642.502.38

t-Wert<.0001*<.0001*0.0003*0.0005*0.0013*0.0146*0.0333*0.0409*0.0487*

Wahrsch.>|t|

Sortierte Parameterschätzer

Parameter Estimates Provide Insights intoCorrelations Between Input Parameters and Outputs

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Leverage Plots of the Statistical ModelIllustrate Actual by Predicted Output Values

0

5

10

15

20

25

30

Pro

tein

Agg

rega

tes

(%) M

easu

red

0 5 10 15 20 25 30Protein Aggregates (%) PredictedP<.0001 r2 =0.99 RMSE=1.295

Diagramm „Beobachtete Werte über Vorhersage“

Prediction formula

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SD = 20% range on every input parameter (worst case) Assumption: normal distribution for input uncertainties Simulation of 10000 batches Aggregate level specification: <25%

No single batch out of specification.

Statistical Models Allow ProcessOutput Simulations Including Input Uncertainties

Spec

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Sound Process CharacterizationHas Significant Benefits for Compliance and Business

Compliance Identification of critical process parameters Interactions between critical variables Justification for operating spaces and acceptance criteria Reproducible quality and yields

Business Improved batch success rates Yield improvements Minimize number of process deviations

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Lonza`s New Approach on Process Characterization Offers Valuable Benefits

Customer Values

A More process knowledge at reduced cost A

B Robust and flexible process B

C Easier process transfer and scale-up C

Compliant Process

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CaC2

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Portsmouth

Singapore

Visp, CH

Slough, UK Kouřim, CZBraine, BE

Porriño, SP

Nansha

Key Injectables

Oral-grade

Our Sites Are Able to Handle YourDevelopment and cGMP Manufacturing Needs

Hopkinton, MA (USA)Microbial U.S. Process R&D GroupSmall to Mid Scale cGMP(150L to 2800L)

Visp (Switzerland)Microbial European Process R&DSmall to Large Scale cGMP(20L to 15,000L)

Kouřim (Czech Republic)Microbial FermentationMid to Large Scale cGMP(75L to 75,000L)

Headquarter in Basel, SwitzerlandEmploys >11,000 people45 major production and R&D facilities

Houston

Houston, TX (USA)Viral-Based U.S. Process R&D GroupMid Scale cGMP(up to 2000L)

Hopkinton

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slide 20www.lonzavirtualtours.com

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Our passion is to deliver sustainablevalue to our customers.

...visit www.lonza.com and meet us at booth 39

Backup Slides

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Lonza’s Interconnected Life-sciencePlatform Comprises Four Divisions

TherapeuticCell Solutions

TestingSolutions

ResearchSolutions

Bioscience

DevelopmentServices

BiologicalManufacturing

ChemicalManufacturing

CustomManufacturing

NutritionIngredients

PerformanceIntermediates

Life ScienceIngredients

Hygiene &Preservation

Water Treatment

MaterialsProtection

Personal Care

Wood Treatment

MicrobialControl

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Cellular TherapeuticsRegenerative medicineTissue engineeringGene therapyViral vectors

VaccinesMicrobialViral

Protein TherapeuticsmAbs / FabsADCsRecombinant proteinsPlasmid DNAEnzymes

Market Focus in Biopharmaceuticals

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Our Microbial Sites Offer Proximity ofR&D, Scale-up and GMP ManufacturingHopkinton, MA

(USA)Visp / Lalden (Switzerland)

Kouřim(Czech Republic)

Employees: ~350 40L to 2x 800L

Employees: ~3000 20L, 50L, 1000L to 2x 15m3

Employees: ~380 15m3, 50m3 and 75m3

Therapeutics and vaccines for multiple indications such as cancer therapy and infectious diseases

Active pharmaceutical ingredients Biopharmaceuticals Antibody drug conjugates (ADC) Peptides and oligonucleotides

Biotransformation Secondary metabolites Recombinant / technical proteins L-Carnitine (CarnipureTM and

CarnikingTM)

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Houston, TX (USA) Lonza Houston, Inc., Houston, TX Employees: 20 Plant/ process:

Process development Scale-up cGMP production Analytical assays Regulatory support

Products: Viral-based therapeutics (viral vector gene

therapy, viral vaccine applications)

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Straindevelopment

and cell banking

Fermentation and recovery

development

Purificationand refold

development

Analytical development

Process validation Support services

Our R&D Services Create a Foundation for Successful cGMP Operations

Lonza Development Services Economically viable processes Robust, scaleable, flexible and reliable processes Broad, full-service offering

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Categories of Customer Projects

Technology Transfer

Process Demonstration

Process Transfer

Technology Transfer

Process Transfer

Technology Transfer

Technology Transfer

Fermentation Development

Fermentation Development

Purification Development

Purification Development

Strain Development

Process Demonstration

Process Demonstration

Process Transfer

Process Transfer

Process Demonstration

Scale-upScale-up

Process Adaptation

Scale-up

High Degree of Process Definition Low

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Permexcis® Virus Production Medium Optimized for use with PER.C6® cell line

Serum-free and chemically defined

Reduced COGs and time to market due to high cell density(> 90%) and viability

Saves validation time No weaning from serum-containing

medium required Minimal lot-to-lot variation

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Broad experience Novel protein therapeutics Antibody fragments Vaccines

(recombinant and attenuated) Cytokines Growth factors

We Have Expertise in All Types of Microbially Derived Products

Interferons Polysaccharide-protein

conjugates Recombinant peptides Plasmid DNA Fusion proteins PEGylated products Products requiring BL2

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Aligned with 7 Markets

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Location 26 miles from Boston

History Founded in 1987 Biopharma CMO since 1997 Lonza acquired Feb 2007

Key productions Ontak®, ATryn® and Increlex®

Our Hopkinton, MA (USA) Site Multiple Capacities to Meet Your Clinical Needs

cGMP Capacities (total volumes) 1 x 150L 1 x 800L 1 x 2,000L 1 x 2,800L(over 500 batches produced since 1998)

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Location 150 km east of Geneva

Key facts Lonza R&D and production

center World’s largest microbial

biopharmaceutical facility dedicated to CMO industry

25-year history in microbial biotechnology

Producing Cimzia®

The Facilities at Our Visp, Switzerland Focus on Clinical and In-market Supply

cGMP capacities (total volumes) 1 x 20L 1 x 50L (new) 1 x 1,000L 2 x 15,000L Additional mid- and large-scale

expansion planning under way

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Lonza founded in Gampel (CH)

Expansion into the US; Foundation of Lonza Inc.

Commissioning of the naphtha cracker in Lalden (CH)

1897 1965 1969 1971 1983

Verbund and Niacin plant in Visp (CH)

Start of the biotechnology research activities in Visp (CH)

MilestonesShort History of Lonza

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Acquisition of the Biotec plant in Kouřim (CZ)

Acquisition of Cell-Tech (Lonza Biologics)

200319961992

First, new multi-purpose plant constructed. Opening of the fine chemicals complex in Visp (CH)

1984

Commission of the SSP (small scale plant) in Visp (CH). Expansion of the production capacities with two new 15m3 plants for HAPIs (highly active pharmaceutical ingredients) in Kouřim (CZ)

MilestonesShort History of Lonza

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Start up of two further 75 m3 biotechnology plants in Kouřim (CZ)Extension of the cGMP mammalian cell culture capacities with three 20’000 liter fermenters in Portsmouth, NH (USA)Start operation of the BPMSS (biopharmaceutical manufacturing small scale) plant with a 1000 liter fermenter and down stream processing at Visp (CH)Sale of Pasadena site, TX (USA)

20052004

Construction of a second Niacinamide plant in China with a capacity of 6000 tons

2006

Acquisition of UCB bio-products peptide business. Braine l’Alleud, (BE)

Larch arabinogalactan acquisition, Cohasset, MN (USA)

MilestonesShort History of Lonza

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Acquisition of Genentech’s mid-scale mammalian biopharmaceutical plant (4 x 10’000 liter) in Porriño (ES)

Commence construction of the first large scale mammalian cell culture facility in Singapore (4 x 20’000L)

2006 20072007

Acquisition of the research bioproducts business and the microbial biopharmaceutical business of Cambrex

Start-up of large-scale microbial manufacturing in Visp (Cimzia for UCB)

MilestonesShort History of Lonza

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Acquisition of amaxa, a technology leader in cell discovery. Cologne (Germany) is the new product development site for the Research Solutions business of Lonza Bioscience.

Acquisition of Algonomics NV (Gent, BE) strengthens Lonza‘s technology offering for biopharmaceutical development.

Joint venture between TEVA & Lonza (TL Biopharmaceutical Ltd) to develop, manufacture and market a portfolio of biosimilars.

Acquisition of Vivante GMP Solutions in Houston, TX (USA) a viral-vaccine and gene therapy company to enter the viral-based manufacturing market.

Acquisition of MODA Technology Partners for paperless quality-control solutions to strengthen the rapid testing solutions platform.

2008 2009 2009 2010

MilestonesShort History of Lonza

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Acquisition of Arch Chemicals Inc. to build the world’s leading microbial control business.

Secondary listing on the Main Board of the Singapore Exchange Securities Trading Limited (“SGX-ST”)

2011 2011 Continued growth…

Milestones 2011