Cambridge Healthtech Institute’s 5th Annual

Antibody Expression
Successful Strategies for Recombinant & Monoclonal Antibody Expression & Optimization
6-7 November 2014

This third part of the protein expression stream for PEGS Europe will focus on the methods and technologies for choosing the correct hosts and achieving the highest possible affinity, yield and effectiveness for both recombinant and monoclonal antibodies for therapeutics and diagnostics.

Thursday, 6 November

12:30 Conference Registration

13:00 Dessert Break in the Exhibit Hall with Poster Viewing


13:30 Chairperson’s Opening Remarks

James Giulianotti, Ph.D., Senior Research Associate, Early Stage Cell Culture, Genentech


CHO Cells and Processes for Antibody Manufacturing: Convoluted History and Insights

Florian WurmFlorian Wurm, Ph.D., Dr. rer. nat., Professor of Biotechnology, Swiss Federal Institute of Technology Lausanne (EPFL)

“Superstar” CHO are THE production system for therapeutic rec-proteins. Sequencing of one CHO cell genome (2013) has extended our knowledge established in the 1960s and 70s. What made CHO cells so popular? The talk will cover the “prehistory” of CHOs as well as key insights and key opportunities that have emerged during the 30 + year biotech history of these cells.

14:05 Studying Design Principles for Bispecific Antibodies: The H-L Pairing Challenge

ItaiBenharItai Benhar, Ph.D., Professor, Department of Molecular Microbiology and Biotechnology, Tel-Aviv University

We present a solution for the correct H-L chain pairing of bispecific IgGs: an engineered disulfide bond between the antibodies’ variable domains. This approach termed disulfide stabilization, replaces the natural disulfide bond between the CH1 and CL domains. By combining Knobs into holes for heavy chains heterodimerization and disulfide stabilization for H-L chain pairing, we efficiently produced several bsAbs in bacteria and in mammalian cells.

14:35 Superior Protein Yields in CHO and HEK-293 Cells Using Novel, Highly Efficient Transfection Reagent

Vjetrovic_JelenaJelena Vjetrovic, Ph.D., Bioproduction Technical Support Specialist, Polyplus-transfection

Low transfection efficiency of CHO cells is a major bottleneck hampering protein production in Transient Gene Expression (TGE). With its 10+ year expertise in transfection, Polyplys-transfection will present superior results obtained in CHO and HEK-293 cells with its novel technically advanced reagent - FectoPRO.

15:05 Refreshment Break in the Exhibit Hall with Poster Viewing 

15:45 Antibody Membrane Switch (AMS) Technology for Facile Cell Line Development

BoYuBo Yu, Ph.D., Co-Founder and CSO, Antibody Research, Larix Bioscience, LLC

Antibody Membrane Switch (AMS) technology is a novel, FACS-based cell line development technology. It utilizes a unique switch mechanism of alternative splicing and site-specific DNA recombinase to turn cells from expressing membrane-anchored antibodies into production cells secreting the antibody. Utilizing AMS technology can reduce cell line screening time from 6-8 months to 2-3 months.

16:15 Bispecific Antibodies in E. coli: It Takes Two to Tango

JamesGiulianottiJames Giulianotti, Ph.D., Senior Research Associate, Early Stage Cell Culture, Genentech

Bispecific antibodies (bisAbs) are being developed by companies in an attempt to address complex disease states. The production of bisAbs in E. coli requires the optimisation of a number of cellular processes within two distinct compartments (cytoplasm and periplasm) and across a single membrane. A number of technologies have been tested and implemented at Genentech that aid in the production of bisAbs in E. coli. This talk will discuss some of our recent work in this area.

16:45 E. coli Periplasmic Expression of Fab’ Fragments

Mark Ellis, Principal Scientist, Protein Expression and Purification, UCB Pharma

This presentation highlights advances we have made in increasing the productive capacity of E.coli periplasmic expression. These are industry leading achievements which suggest that E.coli can maintain a competitive edge over other systems employed in the production of small antibody fragments.

17:15 End of Day One

17:15 Dinner Short Course Registration

17:30 – 20:30 Dinner Short Courses*

*Separate Registration Required. 

Friday, 7 November

08:00 Morning Coffee


08:30 Chairperson’s Remarks

Raphael Levy, Ph.D., Senior Scientist, Preclinical Research and Development, XOMA Corp.

08:35 Application of Cell-Based Assays in Biologics Drug Discovery

LizEnglandLiz England, Scientist 1, Antibody Discovery and Protein Engineering, MedImmune

Cell-based assays can allow identification of therapeutics against complex targets that provide challenges in soluble protein expression such as GPCRs, ion channels and multimeric complexes. In addition therapeutics can be identified with multiple mechanisms of action and screening for function can lead to identification of the right epitope in a shorter time frame. For these reasons cell-based assays have been used extensively for High-Throughput Screening (HTS) within the pharmaceutical industry.

09:05 A Robust High-Throughput Platform to Generate Functional Recombinant Monoclonal Antibodies Using Rabbit B Cells from Peripheral Blood

Stefan Seeber, Ph.D., Principal Scientist, Cell Line and Molecule Development, Roche Pharma Research and Development (pRED)

We have developed a new two-step technology involving the isolation and cloning of single B cells from rabbit blood samples after immunization, screening of the B cell supernatants, followed by automated and high-fidelity B cell PCR of the antibody coding sequences. The antibodies are then directly sequenced and expressed or genetically engineered to create chimeric antibodies as well as bispecific antibodies, antibody fragments and other scaffolds. We will present an example of generating antibodies against a human interleukine receptor.

09:35 Problem Solving Roundtable Discussions

Fine Tuning Cell-Based Assays for Drug Discovery

Moderator: Liz England, Scientist 1, Antibody Discovery and Protein Engineering, MedImmune

  • What do I need to know?
  • Fine tuning the results
  • What options do I have?
  • Can I modify an existing assay?

Solving Bi-Specific Antibody Expression Problems in E. coli

Moderator: James Giulianotti, Ph.D., Senior Research Associate, Early Stage Cell Culture, Genentech

  • What are the challenges going in?
  • Success stories
  • Making better choices

Using the Antibody Membrane Switch to Produce Better Production Cell Lines

Moderator: Bo Yu, Ph.D., Co-Founder and CSO, Antibody Research, Larix Bioscience, LLC

  • How does it work?
  • What hosts can be used?
  • What yield improvements are possible?

10:35 Coffee Break

11:00 Determinants and Impact of Antibody Aggregation on Production and Application

JoostSchymkowitzJoost Schymkowitz, Ph.D., Principal Investigator, VIB Switch Laboratory, KU Leuven

As most proteins, antibodies have a propensity to aggregate that is determined by their primary sequence and aggregation acts as a bottleneck on both production and application. Accurate prediction of antibody quality is currently lacking but would be of value to help identify good antibodies. I will discuss a number of key determinants and how to employ them for this purpose.

11:30 Improved Panning Output and Antibody Fragment Production by Co-Expression with the Peptidyl-ProlylIsomerase, FkpA, in the Cytoplasm of Escherichia coli

RaphaelLevyRaphael Levy, Ph.D., Senior Scientist, Preclinical Research and Development, XOMA Corp.

Bacterial cytoplasmic expression of cytFkpA, a variant of peptidyl-prolylisomeraseFkpA, improved secretion of functional Fab fragments into the periplasm, exceeding the benefits of Fab co-expression with the native periplasmic FkpA. Panning and subsequent screening of large naïve phage antibody libraries in the presence of cytFkpA significantly increased the number of unique clones selected, their functional expression levels, and diversity.

12:00 High-Level Secretion of Recombinant Monomeric Murine and Human Single-Chain Fv Antibodies from Drosophila S2 Cells

FelixRayFelix A. Rey, Ph.D., Head, Départment de Virologie, Unité de Virologie Structurale, Institut Pasteur

We report here a system allowing for easy and efficient cloning of (i) scFvs selected by phage display and (ii) individual heavy and light chain sequences from hybridoma cDNA into expression plasmids engineered for secretion of the recombinant fragment produced in Drosophila S2 cells. The suitability of the produced recombinant fragments for structural studies was demonstrated by crystallization and structure determination of one of the produced scFvs, derived from a broadly neutralizing antibody against the major glycoprotein E2 of the hepatitis C virus.

Pall Life Sciences12:30 Application of Label-Free Technology for Biologics Discovery
Forte Bio
Frances Neal, MSc, Scientist I, MedImmune, LLC. 

ForteBio’s Octet RED384 is a label free system that uses BioLayer Interferometry (BLI) to understand biomolecular interactions in real time. The Octet RED384 system can be applied to evaluate a number of key characteristics of protein interactions to aid biologics discovery. These include: antibody and antibody fragment quantification from crude and purified samples; competition assays for characterisation of receptor – ligand neutralising antibodies or distinguishing different epitopes; and kinetic screens for antibody affinity and antibody fragment off rate ranking. Here we present examples for these different biologics applications, demonstrating the versatility of this system, and its utility in the discovery of biologic therapeutics.

 13:00 Enjoy Lunch on Your Own 


14:00 Chairperson’s Remarks

Elizabeth J. Lamb, Senior Conference Director, Cambridge Healthtech Institute

14:05 Molecular Farming: Recombinant Barley-Produced Antibody for the Detection of the Major Bovine Milk Allergen, β-lactoglobulin

AnneliRitalaAnneli Ritala, Ph.D., Pharm., Docent, Principal Scientist, Project Manager, IPMA C, Plant Biotechnology, VTT Technical Research Centre of Finland

Molecular farming i.e. the utilization of plants and plant cells as production hosts provides a safe and sustainable platform for the production of valuable proteins like recombinant antibodies. In this study, a barley-based production system for beta-lactoglobulin specific IgE antibody (D1 scFv) was developed. The grain-produced D1 scFv was successfully purified using affinity-based chromatographic protocols and its functionality was confirmed. 

14:35 Expression of Single-Chain Variable Fragments Fused with the Fc-Region of Rabbit IgG in Leishmania tarentolae

PeterKirstensenPeter Kristensen, Ph.D., Associate Professor, Department of Engineering, Aarhus University

To our knowledge, this is the first time that antibody fragments with intact Fc-region of immunoglobulin have been produced in L.tarentolae. This system provides an alternative in cases where antibody constructs express poorly in standard prokaryotic systems. Furthermore, in cases where bivalent Fc-fused antibody constructs are needed, using L. tarentolae for expression provides an efficient alternative to mammalian expression.

15:05 T-Cell Dependent Bispecific Antibodies (TDBs)

Teemu Junttila, Ph.D., Antibody Engineering, Genentech, Inc.

TDBs are full-length IgG1 antibody molecules with a long half-life that are able to recruit T cells to selectively eliminate target-expressing tumour cells. TDBs exhibit robust preclinical activity in the treatment of pre-established tumours in immunocompetent transgenic mice. Our studies reveal a general resistance mechanism for T cell recruiting molecules with significant potential as a predictive diagnostic marker and demonstrated that direct polyclonal recruitment of T cell activity together with blockage of T cell inhibitory signaling results in enhanced and durable long-term responses.


15:35 End of PEGS Europe

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