Cambridge Healthtech Institute’s 24th Annual

Recombinant Protein Expression and Production

Maximizing Quantity and Quality while Minimizing Time and Cost

January 18 - 19, 2022 ALL TIMES PST

The utilization of engineered therapeutic proteins for basic research, clinical diagnostics, and therapy continues to expand. Discovery activities and developability assessments require substantial amounts of candidate biomolecules and are challenging to obtain. Thus, an increasing variety of recombinant production expression platforms are being developed. Unfortunately, there is no “universal” production system which can guarantee high yields of recombinant protein, particularly as every biomolecule itself causes its own issues in terms of expression. Hear experts during the Recombinant Protein Expression and Production conference share their insights applying higher-throughput expression and purification as well as developing flexible expression platforms.

Tuesday, January 18

1:00 pm Registration (Sapphire West Foyer)
1:30 pm Refreshment Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)


Session Room: Sapphire 410

2:00 pm Organizer's Welcome Remarks

Mary Ann Brown, Executive Director, Conferences, Cambridge Healthtech Institute

Andrew Hayhurst, Professor, Disease Intervention and Prevention, Texas Biomedical Research Institute

Whether natural spillovers, geographic migrants or unfortunate laboratory escapees, emerging RNA viruses pose grave threats to human health and socioeconomic progress. We combine responsive and forward capable strategies to deliver nanobodies specific to one species or cross-reactive to all species of a viral genus. SARS-CoV-2 was our responsive model used to deliver species specific probes while the genus Ebolavirus was our forward capable model employed for delivering probes that recognize all six species equally. The conserved and polymeric nature of viral nucleoproteins enable even modest nanobodies to be useful long-term molecular probes especially when produced as enzymatic and fluorescent fusions. 

2:40 pm

CyDisCo Production of Functional Recombinant SARS-CoV-2 Spike Receptor Binding Domain

Gloria Borgstahl, PhD, Professor, Biochemistry & Molecular Biology & Pharma Sciences, University of Nebraska Omaha

We developed a bacterial strategy for the expression and purification of a SARS-CoV-2 spike protein receptor binding domain (RBD). Bacterial cytoplasm is reductive and this is problematic when the recombinant protein of interest requires complicated folding and/or processing. The CyDisCo system bypasses this issue by pre-expressing a sulfhydryl oxidase and a disulfide isomerase, allowing the correct folding with disulfide bonds for protein integrity and functionality. We show that it is possible to quickly and inexpensively produce an active RBD in bacteria that is capable of recognizing and binding to the ACE2 receptor as well as antibodies in COVID-19 patient sera.

Diane Retallack, Senior Vice President, Platform Tech & Innovation, Pelican Expression Technology

Rapid and comprehensive strain selection and an early process development program utilizing the Pelican Expression Technology™ were key to establishing the foundation for late-stage success for Rylaze™, a recombinant Erwinia chrysanthemi asparaginase for the treatment of acute lymphoblastic leukemia or lymphoblastic lymphoma in patients who have developed hypersensitivity to E. coli-derived asparaginase. Pelican Expression Technology™, a Pseudomonas fluorescens-based protein expression system is a robust and scalable platform for recombinant protein production.

3:40 pm Refreshment Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)
4:30 pm

Expression of SARS-CoV-2 Surface Glycoprotein RBD in E. coli, and Its Refolding and Purification

Maria Kireeva, Director, R&D, VirIntel, LLC

Cost-effective methods of expression and purification of SARS-CoV-2 spike and its fragments that preserve antigenic properties are essential for development of COVID-19 serology tests. We developed a method of purification of S319-640 fragment containing RBD expressed in E. coli from the inclusion bodies. The antigenic properties of the resulting product are similar to those of the RBD-containing fragment expressed in human cells.


Protein Production Lab Challenges: Methodologies, Strategies, and the Art of Managing Multiple Projects

Panel Moderator:
Richard Altman, Field Application Scientist, Life Science Solutions, Thermo Fisher Scientific

There are many challenges and rewards in operating a protein production lab. This panel will focus on the following topics: ​

  • Lessons learned from managing a protein production workflow during a pandemic.
  • Strategies on how to manage multiple “top priority” projects. Strategies for supporting the professional growth and career development of direct reports.
  • How do we make time for technical development and process optimization?
  • Troubleshooting strategies or how much time should be spent before moving to the next option?
Dominic Esposito, PhD, Director, Protein Sciences, Frederick National Laboratory
Balaji Somasundaram, PhD, Strategy & Operations Manager, Protein Expression Facility, University of Queensland
Elizabeth Stangle, Senior Research Associate, Protein Engineering, Zymeworks Inc.
Bjørn Voldborg, MSc, Head, National Biologics Facility, DTU Bioengineering, Technical University of Denmark
Jessica A. Williamson, PhD, Protein Production Lead, UCB
6:00 pm Close of Day

Wednesday, January 19

7:30 am Registration (Sapphire West Foyer)
8:00 am BuzZ Sessions with Continental Breakfast (Sapphire Foyer)

PepTalk BuzZ Sessions are focused, stimulating discussions in which delegates discuss important and interesting topics related to upstream protein expression and production through downstream scale-up and manufacturing. This is a moderated discussion with brainstorming and interactive problem-solving between scientists from diverse areas who share a common interest in the discussion topic. Continue to check the event website for detailed discussion topics and moderators.

BuzZ Table 6: Multidisciplinary and Inter-institutional Collaborations, How to Succeed?

Bjørn Voldborg, MSc, Head, National Biologics Facility, DTU Bioengineering, Technical University of Denmark
Nathan Lewis, PhD, Associate Professor, Pediatrics and Bioengineering, University of California, San Diego (UCSD)

Harvesting the benefits and overcoming the challenges of collaborations between academic and industrial researchers and institutions specialized in widely spread disciplines and different geographical locations can be demanding but also include unique rewards. ​

  • How to start-up projects
  • Identifying and involving collaborators
  • Managing expectations (IP, Funding, commitment, etc..)
  • Ensuring efficient and smooth collaboration

BuzZ Table 9: Why is it so Difficult to Engineer CHO Cells for Improved Expression and Product Quality?

Rene Hubert, PhD, Director, Biologics Optimization, Amgen Inc.

The full potential of gene editing technologies in CHO cells has yet to be achieved in creating significant improvements in expression and product quality.  Join this discussion group to commiserate, share your experiences, and brainstorm ideas to generate breakthroughs.

  • How do we get the most out of transcriptomics?
  • How do we leverage the secretome and surfaceome in generating improved CHO hosts?
  • What’s the best way to generate and validate candidate genes?​


Session Room: Sapphire 410

9:05 am

Bioengineering RNAs for Targeted Therapy

Aiming Yu, PhD, Professor, Biochemistry & Molecular Medicine, University of California Davis

RNAs have emerged as a novel class of medications with unique mechanisms of actions. Current RNA research and development are restricted to the use of chemo-engineered RNA mimics made in vitro, which are different from natural RNAs produced and folded in vivo. In this talk I will present our novel RNA bioengineering technology for high-yield, large-scale and cost-effective in vivo fermentation production of true biologic RNA agents carrying payload RNAi molecules.

9:35 am

A Novel Recombinant Soluble Complement Receptor 1 Fragment with Enhanced Therapeutic Potential

Matthew Hardy, PhD, Associate Director, Recombinant Proteins, CSL Research

We have generated and characterized CSL040, a recombinant soluble and truncated form of human Complement Receptor 1 (CR1) that potently inhibits all three pathways of the complement system and represents a new and attractive therapeutic candidate for complement-mediated disorders. CSL040 attenuates disease development in multiple animal models, and we demonstrate that sialylation-dependent pharmacokinetics and differential complement pathway inhibition are hallmarks of CSL040 activity in vivo.

Matthew Weinstock, PhD, Chief Technology Officer, Absci

Recent advances in synthetic biology and artificial intelligence promise to dramatically impact the way that pharmaceuticals are developed.  In this presentation I will discuss how Absci is leveraging the power of these technologies to address key pain points in both how protein-based biologics are discovered as well as how cell lines for manufacturing are designed.  

10:35 am Coffee Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)
11:15 am

A Novel Platform to Create Multi-Functional Immunotherapeutic for Cancer

Hing C. Wong, PhD, CEO & Founder, HCW Biologics Inc.

We developed a novel platform (TOBI) centered upon an inert tissue factor scaffold for production of heteromeric and multi-functional fusion protein complexes for therapeutic uses. We will present our characterization of HCW9218 (a bifunctional TGF-ß trap/IL-15 protein complex) and HCW9201 (a fusion complex combining IL-2, IL-15, and IL-18), created based on the TOBI platform, to demonstrate the versatility and potential utilities of these fusion proteins as immunotherapeutic against cancer.

Jennifer A. Maynard, PhD, Henry Beckman Professor, McKetta Department of Chemical Engineering, Cockrell School of Engineering, University of Texas Austin

Fc engineering is gaining interest as a strategy to tune antibody functions but bacterial and mammalian display systems are limited by their inabilities to provide the native glycosylation that supports binding to classical Fc receptors. To address this, we adapted our mammalian CHO display system to engineer human IgG1 Fc for pH-selective FcgRIIIa binding that preferentially activates cellular cytotoxicity at the low pH common to the tumor microenvironment.

Pierre-Alain Girod, PhD, Chief Science Officer, Science, Selexis S.A.

CHO cell volumetric productivity has significantly increased in the past two decades. However, turning these cells into high-producing factories has created an overall metabolic burden. In response to transgene expression pressure, CHO cells undergo phenotypic and genetic changes for continuous improvement.  Selexis’ SUREtechnology Platform™ provides a solution for overcoming the challenges of high productivity tied to metabolic adaptability. 

12:45 pm Session Break
Jiansheng Wu, PhD, Vice President, Protein Sciences, WuXi Biologics

We discuss the challenges in high-throughput protein production for small and large molecule drug discovery. We demonstrate the parameters and design space required to generate high-quality proteins for HTS, antibody discovery, in vivo and developability studies. Supported by our industry-leading platforms, the Protein Sciences department at WuXi Biologics provides production services utilizing various expression systems for the generation of monoclonal, bispecific and multispecific antibodies, and other recombinant proteins.

1:25 pm Refreshment Break in the Exhibit Hall and Last Chance for Poster Viewing (Sapphire Ballroom)


2:15 pm

Chairperson's Remarks

Nathan Lewis, PhD, Associate Professor, Pediatrics and Bioengineering, University of California, San Diego (UCSD)

Lessons Learned from Expression Liabilities in a Four-Chain Bispecific

Rene Hubert, PhD, Director, Biologics Optimization, Amgen Inc.

Multispecific antibodies are increasingly relevant in biotherapeutic pipelines and significant effort exists in engineering the molecules in addition to optimizing vectors, cells, and processes to improve productivity and quality. To understand the causes and effects of chain mispairing impurities in a difficult to express four-chain multispecific hetero-IgG, we investigated consequences of individual and pairwise chain expression. We identified the causes for the low product quality observed from stable cell lines expressing this hetero-IgG and suggest mitigation strategies. This approach provides a general strategy and justification for identifying the molecular and cellular liabilities associated with exceptionally difficult to express multispecific antibodies.

2:50 pm

Elucidating the Host Cell Machinery Supporting Efficient Therapeutic Protein Secretion through Systems and Synthetic Biology

Nathan Lewis, PhD, Associate Professor, Pediatrics and Bioengineering, University of California, San Diego (UCSD)

Each protein secreted by a cell depends on up to hundreds of different host cell proteins for its synthesis, folding, post-translational modification, and transport. However, this secretory pathway machinery required can differ considerably. Using computational modeling and omics data integration, we have developed a pipeline to identify the host machinery that can be augmented to improve secretion of diverse recombinant biotherapeutics of interest. 

3:20 pm

Membrane Mimetics to Facilitate Antibody Screening

Christy A. Thomson, PhD, Principal Scientist, Amgen, Inc.

The ability to identify and characterize therapeutic antibodies targeting multi-pass membrane proteins is hampered by their often-difficult expression and purification.  Historically membrane proteins have been extracted from cell membranes using detergents, however the presence of detergent can hamper many downstream applications. We examined novel methods for membrane protein stabilization, including SMALPs, nanodiscs and amphipols, to isolate a model multi-pass membrane protein and evaluated their utility in lead antibody identification.

3:50 pm

One-Step Engineering the Affinity, Stability and Expression of a Bispecific Molecule

Renhua Ray Huang, PhD, Scientist III, Antibody Engineering, MacroGenics Inc.

Bispecific antibodies are an emerging class of biologics that have shown great promise in cancer treatment. Yet often during preclinical development, many lead molecules fail due to poor biophysical/biochemical properties. Here a phage-display method has been developed, in which engineering of the cyno cross-reactivity, stability and expression of a bispecific molecule was tackled simultaneously, generating multiple lead molecules that showed significant improvement in all three properties.

4:20 pm

Optimization of a Transient Antibody Expression Platform towards High Titer and Efficiency

Elizabeth A. Greene, Scientist IV, Biotherapeutics Molecule Discovery, Boehringer Ingelheim Pharmaceuticals, Inc.

Chinese Hamster Ovary (CHO) cells are one of the major workhorses for Transient Gene Expression (TGE) of recombinant antibodies. Through a matrix evaluation of multiple factors including inoculum, transfection conditions, amount and type of DNA used (including Filler DNA), and post-transfection culture conditions, we arrived at an uniquely optimized TGE process with higher titer and reduced costs and time, thus increasing the overall efficiency of early antibody material supply.

4:50 pm Close of Conference