Cambridge Healthtech Institute’s Nineteenth Annual
Recombinant Protein Expression and Production
Achieving Quality and Quantity
January 10-11, 2017 | Hilton San Diego Bayfront | San Diego, CA
Biopharmaceuticals currently represent the fastest-growing sector of the pharmaceutical industry, driven by a rapid expansion in the manufacture of recombinant protein-based drugs. Consequently, the efficient expression and production of these valuable
biomolecules face challenges in improving their quantity and quality while minimizing time and cost. To meet these demands, an increasing variety of recombinant production 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. To meet the demand, it is crucial to increase the throughput of expression, production
and purification processes and systems.
Cambridge Healthtech Institute’s Recombinant Protein Expression and Production conference explores the newest data and innovations relating to the best choices in hosts/systems, as well as ways to “rescue” existing systems and make
them work more effectively to produce the quality and quantity of the desired biotherapeutic.
TUESDAY, JANUARY 10
1:00 pm Conference Registration
1:30 Refreshment Break in the Exhibit Hall with Poster Viewing
2:00 Chairperson’s Opening Remarks
Donald L. Jarvis, Ph.D., Professor, Molecular Biology, University of Wyoming; President, GlycoBac, LLC
2:05 Harnessing the Power of MS-Based Proteoinformatics for High-Quality Protein Production
Amit Kumar, Ph.D., Graduate of Michael Betenbaugh’s Lab, Johns Hopkins University; Postdoctoral Research Fellow,
Chemical and Molecular Biology, St. Jude Children’s Research Hospital
Hundreds of host cell proteins (HCPs) are produced during the recombinant protein production in CHO cells. These HCPs are a combination of essential proteins for normal cell functioning such as cell growth and non-essential proteins released due to
apoptosis or cell lysis. Regulatory guidelines mandate that HCPs be identified and quantified to ensure patient safety. Here, we present detailed MS-based proteoinformatics methods for studying CHO proteins and elucidating CHO HCP profile.
2:45 The Beginning of the End: HIV-1 Vaccine Design and Production
Jiang Zhu, Ph.D., Assistant Professor, Department of Immunology and Microbial Science, Department of Integrative Structural
and Computational Biology, Scripps Research Institute
The metastability of HIV-1 envelope glycoprotein (Env) has posed a significant challenge to vaccine design and production. We have identified the N-terminus of heptad region 1 (HR1) as the primary cause of metastability and developed an
uncleaved prefusion-optimized (UFO) trimer platform. UFO trimers demonstrated substantially high
yield, purity, and stability, which allowed the multivalent display of gp140 trimer on self-assembling nanoparticles as virus-like particle (VLP) vaccines.
3:15 ExpiCHO: Latest Developments in High-Titer Transient Protein Expression in CHO Cells
Jonathan Zmuda, Director, Cell Biology, Thermo Fisher Scientific
The ExpiCHO™ transient expression system offers a turnkey solution for generating high-titer recombinant proteins for therapeutic drug development, reagent generation and hard-to-express proteins and has become an integral part of the
transient expression workflow in companies around the world. Here, we share the latest data on the ExpiCHO expression system, tips for obtaining maximal performance and applications data supporting protein purification and characterization
as well as protein production up to the multi-liter scale.
3:45 Refreshment Break in the Exhibit Hall with Poster Viewing
4:30 Strategies for Optimizing GPCR Expression and Production: Multi-Milligram Quantities of Functional Cannabinoid Receptor Isolated by Tandem Affinity Chromatography
Alexei Yeliseev, Ph.D., Staff Scientist, Laboratory of Membrane Biochemistry and Biophysics, NIAAA,
Human cannabinoid receptor CB2, a GPCR, is an important target for pharmaceutical drug development. We expressed the functional CB2 receptor and optimized its purification using novel affinity resins StrepTactin XT and EF2 Ca-calbindin-based
resins. Applications of the CDFast chromatographic technique and the results of the SPR binding analysis will be presented as well as examples of selectively stable isotope-labeled CB2 and NMR studies on the agonist- and inverse agonist-bound
5:00 Production of Chemokine/Chemokine Receptor Complexes for Structural Studies
Martin Gustavsson, Ph.D., Staff Scientist, Skaggs School of Pharmacy and
Pharmaceutical Sciences, University of California, San Diego
Chemokine receptors are seven-transmembrane proteins that interact with chemokine ligands to drive cell migration. Despite the development of new methods for expression and purification of seven-transmembrane receptors, production of stable
complexes between chemokine receptors and chemokines remains a challenging task. We present methods for producing purified complexes by co-expression in Sf9 cells. These methods have been successfully used for crystallization and biophysical
experiments of CC as well as CXC receptor/chemokine complexes.
5:30 Close of Day
WEDNESDAY, JANUARY 11
8:00 am Conference Registration and Morning Coffee
8:30 Chairperson’s Remarks
Ashok D. Bandaranayake, Ph.D., Director, Bioprocess Development, Peptide Drug Discovery Initiative, Fred Hutchinson Cancer Research Center
8:35 Optimizing Antibody Expression by Using Natural Framework Diversity and Host Engineering in a Live Bacterial Antibody Display System
Noelle Lombana, Ph.D., Senior Scientific Researcher, Antibody Engineering, Genentech
We present insights into a novel bacterial display system using full-length formats for antibody and antigen in a live cell setting. We discuss ways to improve expression and stability of antibodies in vitro by mimicking the natural antibody selection process, which translates to a mammalian host, as well as ways to improve antibody expression in host expression strains. We also cover novel use of high-throughput screening to study translocation,
stability and protein folding in E. coli.
9:05 Rapid Production of Biologics with Pichia pastoris
Kerry Routenberg Love, Ph.D., Research Associate & Technical Program
Manager, InSCyT Program, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology
Pichia pastoris has demonstrated utility as a host organism, yet relatively little strain engineering has been performed. Here, we present recent results using our genomic and transcriptomic insights to
improve cultivation conditions and strain performance during heterologous protein expression. These upstream process developments have enabled the deployment of Pichia as the production host in a portable
and distributed platform for real-time manufacturing of biologic drugs.
Improved Antibody Quality and Consistency in CHO Cells using a Novel Media Supplement
Adam Elhofy, Ph.D., CSO, Essential Pharmaceuticals LLC
There has been a push to improve consistency and quality of glycolytic patterns. Protein synthesis and post-translational modification occurs on the lipid membranes of the ER and Golgi. Addition of Cell-Ess improves the consistency and
quality of glycosylation while also increasing titer. The use of Cell-Ess resulted in significantly less variation of the glycolytic pattern and increased higher order glycoforms. The novel method of adding lipids results in an improvement
of protein quality and consistency.
10:05 Coffee Break in the Exhibit Hall with Poster Viewing
PEPTIDE AND PROTEIN THERAPEUTICS
10:50 Characteristics and Utility of an Sf-Rhabdovirus-Negative Insect Cell Line for Baculovirus-Mediated Recombinant Protein Production
Donald L. Jarvis, Ph.D., Professor, Molecular Biology, University of Wyoming; President, GlycoBac,
Recent work suggests all Spodoptera frugiperda-derived insect cell lines are contaminated with an adventitious viral agent, now known as Sf-rhabdovirus. This talk focuses on the characteristics of a novel, Sf-rhabdovirus-negative
Sf cell line and its utility as an improved host for recombinant protein production in the baculovirus system.
11:20 A Cell-Free Expression and Purification Platform for Rapid and Flexible Production of Protein Biologics
John Dresios, Ph.D., Chief Scientist and Technical Fellow, Leidos, Inc.
We report on the development of a fluidic process for rapid end-to-end production of recombinant proteins. This process incorporates a bioreactor hosting a cell-free system programmed for transcription/translation of engineered DNA integrated
with a series of configurable downstream purification/formulation modules for process-specific isolation of protein targets. Using this approach, we demonstrate production of two bioactive protein therapeutics, each within 24 hours.
This process is flexible, scalable and amenable to automation.
11:50 Optides: A Novel Mid-Size Medicine Drug Discovery Platform Based on Knotted Proteins
Ashok D. Bandaranayake, Ph.D., Director, Bioprocess Development, Peptide Drug Discovery
Initiative, Fred Hutchinson Cancer Research Center
Knottins are highly disulfide crosslinked peptides associated with the venoms of insects. They inhabit a unique space between antibodies and small molecules but are difficult to make synthetically. We have developed a fully automated mammalian
expression platform to generate these molecules as therapeutic leads. We call these optimized peptides Optides, and our first dye-peptide conjugate, Tumor Paint (Blaze Biosciences), is now in multiple clinical trials as a surgical
aid for resecting tumors.
12:20 pm Selexis SUREscan™: Improving Research Cell Bank Generation & Clonality Verification with Comprehensive Genomic Analysis
Girod, Ph.D., Chief Scientific Officer, Selexis
Using Next-Generation Sequencing technologies combined with proprietary bioinformatics tools (Selexis SUREscan™), Selexis now has the ability to quickly analyze the whole genome of any Selexis-generated research cell bank (RCB). In light of the FDAs recent concerns regarding establishment of clonality for IND and BLA submissions, we will describe how we apply SUREscan™ to improving monoclonality assessment and traceability of RCBs, MCBs and WCBs. Case studies will be included.
12:50 Session Break
1:00 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
2:00 Chairperson’s Remarks
Bjørn Voldborg, MSc, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
2:05 A Predictable, Plug-and-Play Cell Culture Platform Process
James Lambropoulos, MS, Engineer III, Cell Culture Development, Biogen
We have implemented a defined workflow for early stage clinical cell line development. A meta-analysis of several monoclonal antibody products, in multiple host cell lines, demonstrates predictable trends and correlations amongst growth,
metabolite, productivity, and quality attributes. This analysis confirms that our development platform is a robust and reliable workflow for generating representative, high-quality protein material for clinical use, and offers
interesting avenues for further refinement of the process.
2:35 In the Pursuit of High Producers: Use of the Sony SH800 Cell Sorter for the Selection of Cell Lines with Superior Productivity Characteristics
Nadia Amharref, Ph.D., Scientist, Cell Line Development, Vaccine Research Center, National
Institute of Allergy and Infectious Diseases, NIH
Shortening the process of developing and isolating high-producing cell lines remains a challenge. Our study focuses on the development of a simple and rapid method using Fluorescence-Activated Cell Sorting (FACS) for the screening
of high-producing recombinant CHO cell lines. In this method, flow cytometry was partnered with a reporter protein for rapid, early stage identification of clones producing high levels of a therapeutic protein. A cell surface protein
not normally expressed on CHO cells is co-expressed, as a reporter, with the therapeutic protein and detected using a fluorescently labeled antibody. The reporter protein’s expression level accurately predicts the relative
expression level of the therapeutic protein for each clone.
3:05 Fast Cell Line Development for CHO Clones with High-Yield Protein Production Using Euchromatin-Containing BAC Expression Vectors
Anton Bauer, Ph.D., COO, The Antibody Lab GmbH
Upon stable cell line generation, chromosomal integration site of the vector DNA has a major impact on transgene expression. By using chromosomal loci in BACs and random integration into host cell chromosomes, we developed stable high-yield
production cell lines at an unprecedented speed. We performed several case studies for CHO production clones, and we established for antibodies and even difficult-to-express proteins generation of production clones within three
weeks from transfection.
3:35 Refreshment Break in the Exhibit Hall with Poster Viewing
4:30 A Method for Specifically Targeting Two Independent Genomic Integration Sites for Co-Expression of Genes in CHO Cells
Joop van den Heuvel, Ph.D., Research Group Leader, Recombinant Protein Expression, Helmholtz Centre for Infection Research
5:00 Engineering Protein Production Hosts
Bjørn Voldborg, MSc, Director, CHO Cell Line Development, The Novo Nordisk Foundation
Center for Biosustainability, Technical University of Denmark
We are using the combined competencies of scientific groups working within the areas of metabolic modelling, glycobiology, cell line engineering, high-throughput methodology and genome editing tool development to design and engineer
the next generation of recombinant protein production hosts. The most recent results from high-throughput targeted genomic manipulations to engineer cells for tailored and homogenous glycosylation, increased productivity, improved
product quality and more robust bioprocesses will be presented.
6:20-7:20 Reception in the Exhibit Hall with Poster Viewing
7:20 Close of Conference