High-quality proteins are needed both for structural and functional studies, and also to create biological products. Protein expression is the culmination of multistep processes that can be characterized and streamlined to efficiently produce stable biologics. The Protein Expression System Engineering conference reveals engineering strategies for producing viable proteins that are soluble and functional. Speed and yield are critical factors, yet essential steps demand time and resources, including verification and sequence analysis of the gene or protein of interest, codon optimization, vector construction, selecting and optimizing a clone, and selecting a host system.

Learn from Protein Expression experts and see how expression tasks are streamlined for greater quality and productivity.

Recommended Pre-Conference Short Course*

SC9: Overcoming the Challenges of Immunogenicity Assessment

*Separate registration required.



12:35pm  Luncheon in the Exhibit Hall with Poster Viewing

1:40  Chairperson’s Remarks

Jagroop Pandhal, Ph.D., Lecturer, Biological Engineering, Chemical and Biological Engineering, The University of Sheffield


Protein Microarrays for Studies in Biomarkers and Post Translational Modification

Joshua_LaBaerJoshua LaBaer, M.D., Ph.D., Director, Virginia G. Piper Center for Personalized Diagnostics, The Biodesign Institute, Arizona State University

Self-assembling protein microarrays can be used to study protein-protein interactions, protein-drug interactions, search for enzyme substrates, and as tools to search for disease biomarkers. In particular, recent experiments have focused on using these protein microarrays to search for autoantibody responses in cancer patients. These experiments show promise in finding antibody responses that appear in only cancer patients. New methods using click chemistry-based reagents also allow the application of these arrays for discovering new substrates of post translational modification.

2:20  Assisted Design of Antibody and Protein Therapeutics (ADAPT): Recent Advances in the Antibody Affinity Maturation Platform 
Christopher_CorbeilChristopher R. Corbeil, Ph.D., Research Officer, Human Health Therapeutics, National Research Council Canada

To assist affinity maturation of therapeutic antibodies we have developed a platform combining binding affinity predictions with stepwise experimental validation. Starting from the crystal structure of an antibody-antigen complex, an efficient workflow intertwines computational predictions with experimental validation from single-point to quadruple mutants. Examples of employing ADAPT for maturation of multiple antibodies will be presented.

2:50  GlycoExpress: A Toolbox for High Yield Production of Glycooptimized Fully Human Biopharmaceuticals in Perfusion Bioreactors at Different Scales

Steffen Goletz, Ph.D., CEO and CSO, Glycotope GmbH

With the GlycoExpress toolbox, we have generated a set of glycoengineered human cell lines for high yield production and for improvement of the clinical efficacy and side effects of fully human biopharmaceuticals. Independent of the applied cell retention mechanism and production scale, GlycoExpress cells show robust growth and consistent glycosylation along with no measurable differences in product quality between batches, batch sizes, reactor sizes, process control strategies, DSP scales and production site.

A Dual Assay Cell Line for Functional and Internalization Studies
Fen-Fen Lin, M.S., Senior Scientist, Biologics, Amgen, Inc.

3:50  Refreshment Break


4:20  Tiny but Mighty: Harnessing microRNAs for Pathway Engineering of CHO Cell Factories

Simon_FischerSimon Fischer, Ph.D., Scientist, BP Process Development Germany, Boehringer Ingelheim Pharma GmbH & Co. KG

The biopharmaceutical industry is currently facing increasing numbers of new biological entities that often turn out to be difficult to express. These molecules can substantially challenge cell line development in order to establish high-yielding production clones. microRNAs were recently demonstrated as exciting new modulators of cell phenotypes in CHO cells. This presentation provides insights into successful exploitation of microRNAs as next-generation cell engineering tool for CHO production cells.

4:50  Role of Codon Optimization and Signal Peptide towards High Titer in Antibody Production

Saurabh_SenSaurabh Sen, Ph.D., Principal Scientist, Immune Modulation and Biotherapeutics Discovery, Boehringer Ingelheim Pharmaceuticals, Inc.

Multitudes of factors play an integral role towards generation of high antibody titers in a transient expression system. The increasing demands in volume and speed have driven us to explore the need for optimization of the codon bias and use of proper signal peptide during transient expression. The presentation will describe our recent efforts towards this end.
View the Interview

5:20  End of Day

5:15  Registration for Dinner Short Courses

Recommended Dinner Short Course*

SC12: Strategic Bioassay Design and Analysis

*Separate registration required.


8:00 am Registration and Morning Coffee


8:30  Chairperson’s Remarks

Krista Alvin, MS, Associate Principal Scientist, Bioprocess Technology & Expression, Merck, Inc.

8:35  Maximizing the Output from an RBS

Daniel_DaleyDaniel Daley, Ph.D., Associate Professor, Biochemistry & Biophysics, Stockholm University

High-level production of recombinant proteins in E. coli is usually obtained with plasmids containing optimised genetic elements (i.e. promoter / RBS / ori). Despite this, some proteins are still difficult to produce. Our data indicate that incompatibly between the RBS and the CDS is a common cause of poor production. We will present a simple and inexpensive PCR-based method for harmonizing these two elements that boosts production levels considerably.

9:05  A New, Growth Decoupled E. coli Expression System

Gerald_StriednerGerald Striedner, Ph.D., Assistant Professor, Biotechnology, University of Natural Resources and Life Sciences, Vienna

Our E. coli expression system design is strongly focused on making them fit for production conditions. One strategy to improve the capabilities of cells for production of finicky or toxic proteins by decoupling cell growth and product formation. In such decoupled systems, cells allocate full protein synthesis capacity to recombinant protein production.
View Speaker Q&A

9:35  Expanding the E. coli Toolbox: Metabolic Engineering to Improve Protein Glycosylation Efficiency

Jagroop_PandhalJagroop Pandhal, Ph.D., Lecturer, Biological Engineering, Chemical and Biological Engineering, The University of Sheffield

E. coli is a relatively simple cell chassis with no native glyco-machinery and could therefore make uniform homogenous glycoforms. Although attention has been afforded to the different glycans that can be produced, the efficiency of the system is poor (~1-13% of recombinant protein is glycosylated). We work to improve this efficiency, using systematic metabolic engineering, omics-based forward engineering, and inverse metabolic engineering, together with improving quantitative analysis of the proteins using high accuracy mass spectrometry.

10:05 Coffee Break


10:35  CRISPR-Cas9 Mediated Efficient and Complete Knock-In of Destabilization Domain-Tags Allows for Reversible and Regulated Knock-Out of Protein Function

Benhur_LeeBenhur Lee, M.D., Professor and Ward-Coleman Chair, Microbiology, Icahn School of Medicine, Mount Sinai

We present a CRISPR-Cas9 mediated strategy for efficient and complete knock-in of in-frame degron-domain (DD) tags for interrogating the function of genes that are critical for cell growth and viability. Protein levels can be regulated by different small molecules that control the activity of the various degron-domains. CRISPR/Cas9-mediated knock-in of DD tags represents a generalizable and efficient strategy to achieve rapid modulation of protein levels in mammalian cells.

11:05  Overcoming Obstacles to Recombinant Production of Human Homologs of Bacterial Asparaginases Used as Approved Antileukemic Enzymes

Manfred_KonradManfred Konrad, Ph.D., Research Director, Enzyme Biochemistry, Max Planck Institute for Biophysical Chemistry

As an alternative to currently used therapeutic enzymes of bacterial origin, we pursue molecular engineering of human asparaginase homologs. We developed expression strategies to efficiently produce two human enzymes that require posttranslational processing to gain catalytic activity. We show that transient translational pausing due to rare codons may be essential to overcoming protein misfolding and that strong complexation with the chaperonin complex GroEL/ES can be suppressed in order to achieve efficient protein production.

11:35  Chromatin Function Modifying Elements in an Industrial Antibody Production Platform

Mark Ellis, Principal Scientist, UCB-New Medicines

The isolation of stably transfected cell lines for the manufacture of biotherapeutic protein products can be an arduous process. This frequently involves transgene amplification and maintenance over many generations. We assessed four chromatin function modifying elements for their ability to negate chromatin insertion site position effects and their ability to maintain antibody expression. Stability analysis demonstrated that the reduction in expression was mitigated in the clones containing A2UCOE-augmented transgenes.

12:05pm   Cell Engineering to Improve Productivity through High-Throughput Screening Technologies

Krista_AlvinKrista Alvin, MS, Associate Principal Scientist, Bioprocess Technology & Expression, Merck, Inc.

Recombinant protein productivity has increased at an exponential rate over the past 10 years, making the task of further improving mammalian cell production increasingly challenging. Here, we will present the identification of new intracellular targets and the corresponding signaling pathways that affect recombinant protein secretion through high-throughput approaches. It provides a basis for the rational design of both cellular and process engineering strategies to further improve protein production.

12:35 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

1:05 Refreshment Break

Vector, Codon & Clone Engineering

1:35  Chairperson’s Remarks

James Love, Ph.D., Director, Technology Development, Biochemistry, Albert Einstein College of Medicine

1:40  Expression of Multiple Antibody Formats in Mammalian Cells from a Single Phage Display Clone Mediated by RNA Trans-Splicing

Isidro_HotzelIsidro Hotzel, Ph.D., Senior Scientist, Antibody Engineering, Genentech, a member of the Roche Group

Antibody discovery and optimization campaigns usually need different antibody formats such as IgG and Fab fragments for screening, requiring the transfer of inserts in large panels of phage display clones to multiple specialized mammalian expression vectors. We developed a modular protein expression system based on pre-mRNA trans-splicing that bypasses clone reformatting steps and enables the expression of multiple antibody formats in mammalian cells directly from a single phage display clone.

2:10  Optimizing Assembly and Production of Bispecific Antibodies by Codon De-Optimization

Giovanni_MagistrelliGiovanni Magistrelli, Ph.D., Head, Protein Engineering, NovImmune SA

We have developed a native human bispecific antibody format, the κλ body , which relies on the co-expression of three polypeptides, one heavy chain and two light chains. Maximal bispecific antibody production is achieved when expression of the two light chains is relatively equivalent. Of particular significance was the finding that codon de-optimization - instead of optimization - of the chain that is over expressed led to significant improvements in bispecific antibody yield.

2:40  Systematic Approaches to Engineering Antibody and Integral Membrane Protein Expression

James_LoveJames Love, Ph.D., Director, Technology Development, Biochemistry, Albert Einstein College of Medicine

We describe a systematic engineering approach that combined machine learning methods with gene synthesis to explore vector element and codon optimization determinants of protein/antibody expression. Combinations of vector components were designed so that elements are varied systematically and independently; this Design-of-Experiment approach allowed us to sample a large sequence-space without exhaustive testing. We then used advanced machine learning algorithms to assess the contribution of each element to vector performance.

3:10 End of Protein Expression System Engineering

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