Monoclonal Antibody Manufacturing –

What Does it Take to Bring Medicine to Patients

NBC2 miniBIOMAN conference Pinar Cicalese, 12July 2017

Agenda: A Complex Disease

Regulatory Expectations

A First in Class Medicine

A Complex BioProcess

A Global Supply Chain

Collaboration Matters Design, Qualify, Verify

Scale-up Challenges

What is Asthma?

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A respiratory condition marked by spasms in the bronchi of the lungs, causing difficulty in breathing.

It usually results from an allergic reaction or other forms of hypersensitivity.

Allergic Cascade in Asthma is Complex

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Mechanism of Action of Biological Therapies in Asthma

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Severe Asthma Have Social and Economic Consequences

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A First in Class Medicine

• A patient is considered to have severe asthma

when they experience frequent symptoms and

acute exacerbations of asthma requiring

treatment with oral corticosteroids, despite

receiving regular asthma therapy, including

high dose long acting inhaled corticosteroids

• Mepolizumab, a monoclonal antibody, has

been shown to reduce exacerbations by

over 50% in severe refractory asthmatic

patients

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http://www.nucalahcp.com/eosinophil-severe-asthma.html

A Long History of Nucala Process Development/Manufacturing

• Process 1 developed by

GSK 1997-2003

• Manufactured at pilot scale

for clinical studies

2004 2005 2006 2007 2008 2009 2010 2011 2012 2013 2014 2015 2016 2017

• Process 2 developed by GSK

2011-2012

• Manufactured at commercial

scale for Clinical.

• Process 2 PPQ batches

manufactured

• Regulatory Filing / Approval

• Launch in various markets

• Commercial manufacturing

and CPV • Process 1 manufactured at

commercial scale for clinical

studies and PPQ

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Filling, Lyophilisation &

Packaging

Cell Culture &

Purification

Analytical

Testing of both

DS and DP

UK

Nucala Supply Chain is Global

USA

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Waupaca, WI

Veenendaal,

NETHERLANDS Jaffrey, NH

SWEDEN sites:

Uppsala

Bohus

Mjolby IL Sites:

Bedford Park

Downers Grove

Glenview

Pekin

Utica

Waukegan GERMANY Sites:

Darmstadt

Phillipsthal

Waldkraiburg

Logan, UT

JAPAN Sites:

Miyazaki-ken

Oita-ken

Tochigi-ken

Paris, KY

Rittman, OH

Old Fort, OH

NY Sites:

Courtland

Fairport

Grand Island

Oneonta

Rochester

Phillipsburg, NJ

Washington, NJ

Pocomoke City, MD

Sparks, MD

PA Sites:

King of Prussia, PA

Mill Hall, PA

Millersburg, PA

Malvern, PA

Scranton, PA

Souderton, PA

Southampton, PA

Stroudsburg, PA

Whitehall, PA

Wulzeshofen,

AUSTRIA

Kalundborg,

DENMARK

UK sites (see below)

FRANCE Sites:

Molsheim

Aubagne

UK/GB Sites:

Cardiff, UK

Consett, UK

Cramlington, UK

Falmouth, UK

Ilfracombe., UK

PA sites (see below)

St.Louis, MO

…and Complex

over 100 Raw Materials for a bulk drug substance batch

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Fajardo, PR

Yauco, PR

Bruntal, CZECH

Republic Kansas City, KS

Arlington, TX

Midlothian,TX

Oxnard, CA

Elysian, MN

Minneapolis, MN

Geismar, LA

Clearwater, FL

Miami, FL

Danvers, MA

Westborough, MA

Others:

Batesville, IN

Brookfield, CT

Rocky Mount, NC

Green River, WY

Searsport, ME

Typical Process Flow for a Mammmalian Protein Drug

Substance

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Affinity

Viral Inactivation (Low pH Treatment)

Anion Exchange

Hydrophobic Interaction

Bulk Drug Substance

Typical Process Flow for Lyophilized Drug Product

Received Bulk

stored frozen

Shipment to Distribution

Point

Labelling & Packaging

Store in Quarantine

for Release Testing

DRUG PRODUCT CQA’s

Sterility

Container Closure Integrity

Particulate Matter

Residual Moisture

Reconstitution Time

Content Uniformity

Visual Appearance

Volume Delivered

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Scale-up Challenges for Bioreactors

Up to 1000 fold scale difference between model and commercial equipment

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Small changes in the cell’s environment (O2, temp, pH, nutrients) can change the relative ratio

of variants, or even produce new variants.

• There is no 1st principles model for this

• Only empirical models exist

Small changes to process can result in differences in product impurity, glycoform, quality

attributes

Leading to changes in:

• Potency/efficacy/stability

• Immunogenicity

• Pharmacodynamics/pharmacokinetics

Scale-up Challenges for Chromatography

Linear scale-up (little economy-of-scale), expensive resin!

Process Scale: 100 L 1,000 L 10,000 L

Product @ 1 g/L: 100 g 1 kg 10 kg

Binding Capacity: 25 g/L resin 25 g/L resin 25 g/L resin

Bed height: 20 cm 20cm 20 cm

adapted from André Dumetz

2m ID

(630L-20cm packed bed)

$5MM per resin pack!

45cm ID 1cm ID

14cm ID

only lasts a few yrs

before it must

be replaced!

Resin cost: 10,000$/L 10,000$/L 10,000$/L

Packed Column: $40,000 $200,000 $1,500,000

Leads to back

pressure buildup

Column diameter: 15 cm 45cm 100 cm

Cycles : 1/ batch 2/ batch 3/ batch

Resin volume: 4 L 20 L 150 L

Linear velocity: 300 cm/hr 300 cm/hr 300 cm/hr

Flow rate: 1 L/min 2 L/min 8 L/min

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Example

R&D and Manufacturing Must Collaborate

R&D knows best:

1. Process Requirements.

2. Analytical methods and

product quality attributes.

3. Scale-up experience from

laboratory to pilot scale.

4. Development history and

improvement options.

5. Have Knowledge and

Severity of risks on product

quality and safety.

Manufacturing knows best:

1. Equipment capability & facility

limitations.

2. Operational constraints.

3. Scale-up experience from pilot

to commercial scale.

4. Local environmental and

governmental regulations

5. Can assess Occurrence and

Detectability of risks.

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Overview example of a process validation lifecycle:

Design, Qualify, Verify

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Process

Development Small Scale Model

Qualification

Proven Acceptable

Ranges (PAR)

Studies

Design Qualify Verify

Small Scale

Full Scale

Scale up and

process

“shakedown”

GMP batches,

including equipment /

cleaning validation

Process

consistency,

BLA enabling

data packageCommercial stockbuild and

product lifecycle management

File for

License

Eng

BatchesClinical

III

supply

Batches

PPQ

Batches

Build Commercial

Launch Stock

Regulatory

Agency Approval

Ongoing

Commercial Supply

opTRA & Periodic review

Initial

control

strategy

Continued process verification

and shelf-life extension stability

CPV plan

protocol

R&D

Donor

PTD

Commercial site

receiver PDC

reference

standard

Development of batch control documents and automation:

SOPs, sample plans, batch records, and specifications

Commercial

control

strategy

Monitor and gather long term

process performance feedback

for future projects

Small Scale

Validations

Ancillary

Qualifications

Update control

strategy as needed

Pre-Approval

Inspection

Essential Terms: Critical Quality Attributes

and Critical Process Parameters

Control for Key OUTPUTS:

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Control of Key INPUTS :

A Critical Process Parameter (CPP) is a process

parameter (fixed or adjustable variable) whose variability

has an impact on a critical quality attribute and therefore

should be monitored or controlled to ensure the process

produces the desired quality. (ICH Q8)

A Critical Quality Attribute (CQA) is a physical, chemical,

biological, or microbiological property or characteristic that

should be within an appropriate limit, range, or distribution

to ensure the desired product quality. (ICH Q11)

ICH-ALIGNED TERMINOLOGY WHAT THIS MEANS IN PRACTICE

Define the quality, safety, and efficacy targets related

to route of administration, dose form, bioavailability,

and strength of the drug.

Determine Drug Product CQAs

Select Formulation, Presentation, and Process

Confirm CQAs and Analyze Failure Mode Effects

Link material attributes & process parameters to CQAs

Determine Design Space and a control strategy

Identify CPPs

Prove acceptability of setpoint ranges

Set Testing Strategy (in-process, release, stability)

Process Validation

Manage changes and continued improvement

Manage residual risks with growing experience

Maintain a state of control

Quality by Design Approach to Process Development

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Quality Target Product Profile (QTPP)

Design Selection

Lifecycle

Management

Control Strategy

PARs &

Design Space CQAs

Risk Assessment

PPQ is not the End, It’s Just a Beginning of a Cycle!

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Potential Improvement

Identified

Risk Assessment and Change

Management

Commercial Control Strategy

Data Monitoring, and Trending

Analysis

On-going Verification of Effectiveness

Periodic review and gains in

process understanding

Process Development

Proposed Process Requirements

& Controls

Process Performance Qualification

R&D determined critical quality attributes

for product safety, efficacy, and dosing,

and product testing rationale

Designated CPPs and Risks

Parameter/Attribute Relationships

Operational Control Risks

Raw Material Risks

Focus on

means & methods

to detect and

act on the

state of control

Lab Scale Study Plan of Characterization & Validation

Plan Product Validation Master Plan

Cell Bank Characterization

In vitro cell age characterization

Validate Clearance of

Host Related Impurities,

Process Related Residuals

Cell Culture

Bioreactor PARs

Bioreactor

Scale-down Model

Validate Lifetime of

Packed Chromatography Resins

& set of UFDF Membranes

Purification Process

Scale-down Model

Assess risk for virus entry via raw materials.

Validate Viral clearance capacity

In-process Leachables from

polymers w/product contact

Purification

Process PARs

Validation of Process Intermediate Stability

Validate

Shipment Routes

Freeze/Thaw Stability,

Shake Stability,

Container Closure Integrity

R&D DS Characterization Studies R&D DS Validation Studies

Acceptable Hold points, Times, &

Conditions for Process Intermediates

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• Process Performance Qualification (PPQ) including Cell Culture and

Purification (Bulk Drug Substance) including Reprocessing

• Media Pre-Filtration Time Allowance & Shelf-life Storage Expiry

• Demonstration of Seed Lab & Bioreactor Aseptic Conditions

• In Vitro Cell Age (IVCA) qualification at scale

• Buffer Formulation Control Limits

• Microbial Control Alert Limits and Specifications (Bioburden &

Endotoxin)

• Chromatography Column Resin Lifetime at scale

• UF/DF Membrane Filter Lifetime at scale

• BDS Shipping Qualification

• Process Performance Qualification (Drug Product)

Major Studies Performed in the Manufacturing Facility

to Support Process Validation

Note: Pre-requisites are Facility, Utility, Environment, Equipment, Sterilization, Cleaning, Method,

Instrument, Computer Control/ Historian Validations

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How Much Work Does it Take to Perform PPQ?

• Variables - total of 296 tracked

• Less Compared to more

than 7500 for clinical

• Samples - total of 2,571

• Reports – 150+ pages

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Stages A CQA PP CPP

1-2 30 2 48 0

3-4 15 9 14 8

5-12 25 72 19 54

Key Learnings and Recommendations

1. Use previous process knowledge (leveraging of previous PPQ data)

2. Document all rationale in PPQ protocol (seed train, back-up thaws, maintenance

steps)

3. Use control strategy as your ally (300 vs >7500 variables in clinical)

4. Unexpected things happen, keep calm, look forward

1. Investigations

2. Chromatography changes

3. Inclement weather, facility shutdown

5. Pay attention to sample tracking (more than 2500 samples, none missed!)

6. Brace and plan for data entry and verification (employed temp workers)

7. Learn from other projects

1. Undiscovered sources of process variability (β-glucans)

2. Acidic CO2 permeation (dry ice)

8. Have regulatory filing in mind

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Agency Interactions During the First Year of Nucala Lifecycle

> 30 Interactions with multiple agencies from Late 2014 to Late 2015

> 320 Regulatory Questions

Key areas of focus:

1. Microbial Control

2. Proven Acceptable Ranges

3. Hold Steps

4. BDS Shipping and Shelf Life

5. Reprocessing Justification Clarification - 1X at Scale vs 3X Small Scale

6. Agency Specific Requests for Additions to Specifications

7. Proactive Filing of Investigations and CAPAs

8. Prepared Backup Responses for Fast Turnaround

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"Success" by Ralph Waldo Emerson

What is Success?

To laugh often and much;

To win the respect of intelligent people

and the affection of children;

To earn the appreciation of honest critics

and endure the betrayal of false friends;

To appreciate beauty;

To find the best in others;

To leave the world a bit better, whether by

a healthy child, a garden patch

or a redeemed social condition;

To know even one life has breathed

easier because you have lived;

This is to have succeeded.

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Everything we do is to ensure they can

do more, feel better, live longerEach DS batch makes more than 10,000 vials.

Each DS batch helps over 750 patients for a year

Any questions?

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Pinar O. Cicalese Global Product Owner, GSK Global Manufacturing and Supply

NBC2 Advisory Board Member

• Pinar held various technical and management roles at GSK, EMD Pharmaceuticals, and Sequel

Genetics during her over 15 years career in the biopharmaceutical industry. Her experience

encompasses early R&D, late stage development and commercial manufacturing support of

bioprocesses. Her skills span bioprocess engineering, molecular biology and technology

transfer of mammalian and insect cell cultures. Pinar has led scale-up and process transfers

for clinical and commercial manufacturing processes using batch, fed-batch, and perfusion

technologies. She has a proven track record of managing technical teams for process

development, new product launches and validated products with successful process

improvements.

• In her current role as Global Product Owner, Pinar provides leadership to various business units

by ensuring process understanding, control, and capability is maintained over a product’s global

supply chain. She is responsible for management of changes and potential risks via cross-

functional planning, negotiation and influence at all levels of the organization.

• Earlier in her career, Pinar has been an Assistant Professor in Molecular Biology and Genetics

Department, and taught various Chemical Engineering and Molecular Biology related courses

in a variety of universities.

• Pinar has a Ph.D. in Chemical Engineering from Drexel University where she was recipient of

Mollye and Mitchel Glick Scholarship and The Merck Scholar Award . Pinar also was a Fulbright

Scholar during her Ph.D. She has a Master in Science and Bachelor in Science degrees in

Chemical Engineering from Bogazici University in Turkey.

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