Thomas Sterovsky - Polymun Scientific (Vienna Conference, April 2016)

Polymun Scientific Immunbiologische Forschung GmbH

Monoclonality - Challenge or Chance Thomas Sterovsky

Developing and Manufacturing Biopharmaceuticals

and Liposomal Formulations for Human Application

Donaustraße 99, 3400 Klosterneuburg, Austria

Polymun Scientific Immunbiologische Forschung GmbH

Inspected by EMA (June 2015) and FDA (October 2013)

CONTRACT DEVELOPMENT OF BIOPROCESSES

for the production of bio-pharmaceuticals for human application with focus on

mammalian cell culture products

CONTRACT MANUFACTURING OF BIOPHARMACEUTICALS

production license according §63 of the Austrian pharmaceutical law

LIPOSOMAL FORMULATION OF DRUGS AND VACCINES

formulation development and production of GMP-material

RESEARCH REAGENTS

manufacturing and distribution, mainly HIV reagents and recombinant trypsin

OWN R&D PROJECTS

funded by revenues from contract development and contract manufacturing

Core Activities

Reference Projects Biopharmaceuticals

Baxter AG

manufacturing of monoclonal antibody for affinity purification of Protein C

GeNeuro SA

process development and manufacturing of humanized antibody GNbAC1

for treatment of multiple sclerosis

GlaxoSmithKline

process development and manufacturing of recombinant human soluble

ACE2 for clinical studies

Imperial College London

process development and manufacturing of recombinant HIV envelope

protein for vaccination studies

Polymun´s Own Product Pipeline

BIOSIMILARS

rhFSH: validated production process at Polymun, market authorisation in

Europe, registration ongoing in several other countries and regions

worldwide

Epo: validated production process at Polymun, out-licensed for

USA/Canada (phase III study ongoing)

LIPOSOMAL SOD – LIPOXYSANTM

Clinical studies

RECOMBINANT HUMAN AND PORCINE TRYPSIN

Process enzyme, cell culture enzyme

EAVI Horizon 2020

EAVI2020

Financed by the European Commison, the European AIDS Vaccine

Initiative brings together leading HIV researchers from public

organisations and biotech companies from across Europe, Australia,

Canada and the USA in a focused effort to develop protective and

therapeutic HIV vaccines.

The EAVI2020 consortium, which is led by Imperial College London,

unites scientists from 22 institutions, pooling their knowledge and

expertise to develop novel candidate vaccines that can be taken through

to human trials within five years. EAVI2020 is funded with an EU-grant

under the health program of Horizon 2020 for research and innovation.

EAVI Horizon 2020

GP140

Envelope glycoprotein of HIV (Env)

Truncated form of gp160

Image credit: US National Institute of Health

Tommi A. White et al.

J. Virol. 2011;85:12114-

12123

EAVI Horizon 2020

POLYMUN WORK PACKAGE

Production of GMP grade material of eight different variants of gp140

Production of GMP grade material of a h-mAb (PGT145)

CRITICAL ISSUES

Time line – fast and reliable/robust cell line development

Expression of stable and fully cleaved gp140 trimers

Fast transfer to GMP level

Scalable production system

Work Flow Optimization

Jayapal, K P., et al. Chemical Engineering Progress 103.10 (2007): 40.

Work Flow Optimization

POINTS TO CONSIDER

Host cell line evaluation

Genetic construct

Work flow (cloning/screening)

Media optimization

Process development

Regulatory requirements

Work Flow Optimization

GENETIC CONSTRUCT

~220

kbp

<10 kbp

Rosa 26 BAC

↑ Transcriptional efficiency

↑ Specific productivity

↓ Chromatin positional effects

• Many genes on one BAC possible

• No Gene amplification necessary

• Less screening effort

Common Vector

Work Flow Optimization

WORK FLOW

Transfection Selection Amplification

MCB Process

Development

Cloning

Protein

productivity

assessment Clone Selection

One step single cell

cloning supported by

imaging system

Imaging System

Clonality Confirmation

Cell Monitoring

Focus Assurance

Quality Imaging

Speed

Certainty

Reporting

Single cell confirmation image on Day=0

Cell is not stuck to another cell

Monitor cell division, viability and growth over

next few days to form single colony

100% focus assurance across the microplate

allows for qualification of your cloning method

Highest quality brightfield imaging required for

the routine clone screens

Rapid imaging, image capture and clonal

reporting allows for single round of cloning

Provides photo proof of clonally derived cell line

Clonality Report can be used as part of filing

package with regulator

Imaging System

Day 0 Day 1 Day4/5 Day10/14

Review wells, select the wells that are clonal

Imaging System

Day 0 Day 1

Imaging System

Step 1

Select Data

Step 2

Generate Report

Step 3

Distribute

Proof of Concept – Historical Data

CN54GP140 AS MODEL PROTEIN

Cell line established in the

mid 90s at Imperial College of

London

GMP – production for clinical

material

DNA sequence was cloned into a BAC vector

CN54gp140 perfect model protein to evaluate new cell line

development setting for EAVI requirements

Proof of Concept

CN54GP140 AS MODEL PROTEIN

Lead clone within 12 weeks

after transfection

Proven clonality

Product demand can be

covered easily

Day 1 Day 2 Day 12 Day 5

Summary – Monoclonality Challenge AND Chance

BENEFITS OF THE ESTABLISHED SETTING

Time requirement was reduced from 20-30 weeks to 12 weeks

Productivity is easily feasible for project demand. (without the

need of further process development at this project stage)

Evaluation of product quality in early stage of the project

IMAGING SYSTEM

• Confirmation of clonality

• Simple implementation of relevant data in

different reports

• Additional information during clone selection

Thank you www.polymun.com