2014 Archived Content

Difficult to Express Proteins

Day 1 | Day 2 | Download Brochure | Speaker Bios 



Antibody Humanization via One Hot Homology Model (Hands-On) Workshop - View Detailed Agenda 

In silico Immunogenicity Predictions (Hands-On) Workshop - View Detailed Agenda 

*Separate registration required

7:00 am Registration and Morning Coffee

» Plenary Keynote Session

8:30 Chairperson’s Opening Plenary Remarks

Kristi Sarno, Chair, Greater Boston Chapter, Women in Bio; Director, Business Development, Pfenex, Inc.

8:40 Harnessing the Patient’s Immune System to Combat Cancer

Peter CR Emtage, Ph.D., Vice President, Immune Mediated Therapies, MedImmune

With recent FDA approvals, modulation of the immune system is now a clinically validated approach in the treatment of some cancers. At MedImmune, the Oncology Department is developing assets and expertise in Immune Mediated Therapy of Cancer (IMT-C). The challenges from a drug development perspective are multi-fold. The talk will focus on the relevance of preclinical models and translational science to address key issues, including dose selection and rationale combinations.

9:25 Building Regeneron’s Pipeline: From Trap Technology to the VelocImmune Platform to Veloci-Next

George D. Yancopoulos, M.D., Ph.D., President, Regeneron Laboratories; CSO, Regeneron Pharmaceuticals, Inc.

George D. Yancopoulos, M.D., Ph.D., who is the Founding Scientist, President, Research Laboratories and Chief Scientific Officer of Regeneron Pharmaceuticals, one of the world’s top biotechnology companies, will discuss how he and his colleagues exploited a commitment to science and technology to start the company, withstand years of challenges and failures, and emerge with a pipeline of promising technologies and novel/biologics that are beginning to bring hope to countless patients and their families.

10:10 Grand Opening Coffee Break in the Exhibit Hall with Poster Viewing

Using The Science To Your Advantage 

11:05 Chairperson's Remarks

Benjamin Doranz, Ph.D., CSO, Integral Molecular, Inc.


11:10 Construct and Expression Optimization of the G Protein-Coupled Neurotensin Receptor for Structure Determination

Reinhard GrisshammerReinhard Grisshammer, Ph.D., Biotechnology Core Laboratory, National Institute of Diabetes and Digestive and Kidney Diseases

In the past few years, we started to see an explosion in the field of GPCR structure determination with structures of more than 20 unique receptors now deposited in the Protein Data Bank. This exciting progress required a tremendous amount of methods development contributed by many laboratories. I will discuss our approaches that led to successful overproduction, crystallization and structure determination of the neurotensin receptor NTS1 bound to its peptide agonist.

11:40 Artificial Environments and Quality Modulation of Cell-Free Expressed GPCRs: Case Studies of the Human Endothelin and Gonadotropin Releasing Hormone Receptors

Erika OrbánErika Orbán, Ph.D., Postdoctoral Fellow, Institute of Biophysical Chemistry, Goethe University

Protocol development and defining the optimal biosynthetic environment for functionally folded and stable membrane protein samples is a complex issue and requires systematic evaluation of numerous additives. We present case studies for the optimization of cell-free expression protocols of the human Endothelin and Gonadotropin Releasing Hormone receptors. The co- and post-translational solubilization of the synthesized GPCRs in several hydrophobic environments including nanodiscs, amphipols and surfactants was analyzed.

12:10 pm Discovery of MAbs Against Difficult GPCRs, Ion Channels, and Transporters

Doranz_BenBenjamin Doranz, Ph.D., CSO, Integral Molecular, Inc.

To enable the isolation, characterization, and engineering of MAbs against challenging membrane protein targets, Integral Molecular has developed the MPS Discovery Engine™ platform, encompassing Lipoparticles for concentrating native membrane proteins and Shotgun Mutagenesis for membrane protein engineering and epitope mapping. Using the MPS platform, we have generated inhibitory MAbs against the ion channel P2X3 for treating neuropathic and inflammatory pain, and have ongoing discovery programs against additional GPCR, ion channel, and transporter targets.

12:40 Luncheon Presentations (Sponsorship Opportunities Available) or Lunch on Your Own

1:40 Session Break

Novel Technologies to “Get It Done” 

2:00 Chairperson's Remarks

2:05 TAPBOOST Technology: Novel Technology to Enhance the Production of Hard-to-Produce Therapeutic Recombinant Proteins

Akinori HishiyaAkinori Hishiya, Ph.D., Director, Biology, Boston Strategics Corporation

A proprietary protein (TAPBOOSTER) is expressed together with the therapeutic protein. The TAPBOOSTER protein binds to targeted proteins and enhances its production specifically. TAPBOOSTER has successfully enhanced the production of many therapeutic recombinant proteins including monoclonal antibodies and Fc fusion proteins. The technology is particularly effective in enhancing the production of proteins where incorrect folding may result.

2:35 Rapid Screening of Membrane Protein Expression in Transiently Transfected Insect Cells

Hao ChenHao Chen, Ph.D., Sr. Scientist, Protein Technologies, Amgen, Inc.

Membrane proteins play critical roles in many biological processes and constitute the majority of all drug targets. However, producing correctly folded and stable membrane proteins is often difficult and requires intensive protein engineering and optimization. Here, we present a rapid method for screening membrane protein expression in insect cells.

3:05 Cell-Free Systems: Functional Modules for Synthetic and Chemical Biology

Marlitt StechMarlitt Stech, MSc, Cell-Free Protein Synthesis, Fraunhofer IBMT-Potsdam

Here we present recent advances for the synthesis of glycoproteins, proteins containing disulfide bridges, membrane proteins, and fluorescently labeled proteins. The basis for the expression of these difficult-to-express target proteins is a translationally active cell extract which can be prepared from eukaryotic cell lines such as Spodoptera frugiperda 21 (Sf21) and Chinese hamster ovary (CHO) cells.

3:35 High-Yield Membrane Protein Expression from E. coli Using an Engineered Outer Membrane Protein F Fusion

Bryan BergerBryan Berger, Ph.D., Head, Berger Lab; Assistant Professor, Chemical Engineering, Lehigh University

An Escherichia coli-based membrane protein overexpression system utilizing an engineered bacterial outer membrane protein F (pOmpF) fusion has been developed. Full-length human receptor activity-modifying protein 1 (RAMP1) was expressed using pOmpF, solubilized in FC15 and purified to homogeneity. Our approach provides a useful, complementary approach to achieve high-yield, full-length membrane protein overexpression for biophysical studies.

4:05 Refreshment Break in the Exhibit Hall with Poster Viewing

4:45 Problem Solving Breakout Discussions

Never mind the PTMs, What about the folding?

David O’ConnellrModerator: David O’Connell, Ph.D., School of Biomolecular and Biomedical Research, University College, Dublin; Conway Institute of Biomolecular & Biomedical Research, UCD, Dublin

  • Does overexpression of mammalian proteins in lower eukaryotes really fit the bill
  • Where heterodimers are concerned, how can we be sure of correct folding to promote complex formation
  • Apart from posttranslational modifications how confident can we be of structural integrity
  • What are the key assays to determine proper folding
  • What assays can we develop in the discovery lab e.g, binding analysis, protein-protein interaction
  • When is overexpression just too much?

Overcoming Problems with Membrane Protein Characterization

Bryan BergerModerator: Bryan Berger, Ph.D., Head, Berger Lab; Assistant Professor, Chemical Engineering, Lehigh University

  • Alternative techniques to improve stability during purification (protein engineering, fusion proteins)
  • Assays to characterize co-receptor complex formation and stability
  • Balance between inclusion body and membrane integration during overexpression to improve yield
  • Correlating detergent and lipid properties to membrane protein stability during purification

Effects of Preliminary Data on Protein Expression Strategy

Brian ChiswellModerator: Brian Chiswell, Ph.D., Owner and Founder, Dimesions BioSciences

  • Which data is most influential in deciding to proceed?
  • Which data brings the project to a halt?
  • When should optimization begin?
  • Should multiple constructs be carried through entire characterization and scale-up?

Recombinant Expression of Toxins and Peptides

Jacques DumasModerator: Jacques Dumas, Ph.D., Head of Protein Production Vitry, Biologics SCP, Vitry Research Center, sanofi

  • Expression systems
  • Peptide size limitations
  • Mammalian versus microbial
  • Is recombinant expression competitive to synthesis

Membrane Protein Expression in Insect Cells

Hao ChenModerator: Hao Chen, Ph.D., Scientist, Molecular Structure, Amgen

  • Advantages/disadvantages of using insect cells as the expression host
  • Expression vectors and construct design
  • New developments

5:45 Welcome Reception in the Exhibit Hall with Poster Viewing

6:45 End of Day

Day 1 | Day 2 | Download Brochure | Speaker Bios 

Japan-Flag Korea-Flag China-Simplified-Flag China-Traditional-Flag