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Fifteenth Annual
Choosing, Designing and Optimizing Hosts and Platforms 
Methods, Strategies & Applications 
January 24-25, 2013 
 

  

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FRIDAY, JANUARY 25

7:15 am Breakfast Presentation (Sponsorship Opportunity Available) or Morning Coffee

 

Engineering for High-Throughput and Enhanced Performance 

8:15 Chairperson’s Remarks

David K. Karig, Ph.D., Senior Professional Staff, Research and Exploratory Development Department, Johns Hopkins Univeristy Applied Physics Lab 

8:20 Algae-Produced Malaria Transmission Blocking Vaccines    

James Gregory, Ph.D., Researcher, Division of Biological Sciences and the San Diego Center for Algae Biotechnology, University of California San Diego

Here we demonstrate that algae are the first recombinant system to successfully produce an unmodified and aglycosylated version of Pfs25 or Pfs28. Thus, algae are promising organisms for producing cysteine-disulfide-containing malaria transmission blocking vaccine candidate proteins.

8:50 Protein Glycosylation Systems in Bacteria and Their Application in Vaccinology and Diagnostics

Mario F. Feldman, Ph.D., Alberta Glycomics Center, Department of Biological Sciences, University of Alberta

Immune responses directed towards surface polysaccharides conjugated to proteins are effective in preventing colonization and infection of bacterial pathogens. Presently, the production of these conjugate vaccines requires intricate synthetic chemistry for obtaining, activating, and attaching the polysaccharides to protein carriers. Glycoproteins generated by engineering bacterial glycosylation machineries have been proposed to be a viable alternative to traditional conjugation methods.

9:20 Ninety Six-Well Plate as a Scale-Down Model for Multi-Parallel High-Throughput Production Clone Selection

Yung-Shyeng Tsao, Ph.D., Senior Principal Scientist, Protein Expression Technologies, Biological Process Development, Merck Research Laboratory

In bioprocess development, the 96-well plate format has been widely used for high-throughput screening of production cell line or culture conditions. We have employed Fluorinert, a perfluorocarbon, to improve oxygen transfer in 96-well plate and to enable the growth of a mammalian cell line expressing a recombinant monoclonal antibody to match the performance in shake flasks.

Pfenex Inc.9:50 Protein Expression Enabled by Pfenex Expression Technology™

Hongfan Jin, Ph.D., Senior Scientist, Molecular Biology,  Pfenex, Inc.

Availability of active protein is critical in every product development program. Leveraging the high-throughput Pfenex Expression Technology platform, milligram amounts of purified proteins can be produced in support of early stage in vitro and in vivo evaluation.  Case studies will provide an overview regarding how Pfenex scientists accomplish this task.

10:05 Coffee Break in the Exhibit Hall with Poster Awards

11:00 Expression Optimization and Synthetic Gene Networks in Cell-Free Systems

David K. Karig, Ph.D., Senior Professional Staff, Research and Exploratory Development Department, Johns Hopkins Univeristy Applied Physics Lab

Most efforts in synthetic biology focus on engineering living cells, yet cell-free systems offer simpler and more flexible contexts for many applications. We characterize repressible T7 promoters, transcriptional terminators, ribosome binding sites, and reporters in a cell-free context. We then explore approaches for composing regulatory systems and implement inducible negative feedback in E. coli extracts and in the PURE system.

11:30 MOPED: Model Organism Protein Expression Database

Eugene Kolker, Ph.D., Bioinformatics and High-Throughput Analysis Laboratory, Seattle Children’s Research Institute

The Model Organism Protein Expression Database (MOPED, htttp://moped.proteinspire.org) is a new and expanding proteomics resource that enables rapid browsing of protein expression information from publicly available studies on humans and model organisms. MOPED is designed to simplify the comparison and sharing of proteomics data for the greater research community.

12:00 pm Strain Engineering for Enhanced Accumulation of Recombinant Membrane Proteins in Escherichia coli

Georgios Skretas, Ph.D., Institute of Biology, Medicinal Chemistry and Biotechnology, National Hellenic Research Foundation, Greece

Low yields of recombinant expression represent a major barrier to the physical characterization of membrane proteins. To address this issue, we have generated engineered E. coli strains that accumulate enhanced amounts of recombinant membrane protein. We will describe the isolation of these strains through the creation of libraries of genetically modified bacteria, coupled with the use of appropriate high-throughput expression screens for the identification of the desired high-expression clones.

CEVEC 12:30 CAP and CAP-T Cells: Production of Therapeutic Proteins, Antibodies and Vaccines in a Human Expression Platform Hartmut Tintrup, Ph.D., Vice President, Business Development & Licensing, CEVEC PharmaceuticalsHuman CAP (CEVEC's Amniocyte Production) and CAP-T  cells allow for stable and transient production of various biotherapeutics while maintaining authentic human posttranslational modifications. The versatile cells exhibit exceptional yields for antibodies and complex proteins and are also an excellent tool for vaccine/virus production.

Catalent12:45 LUNCHEON PRESENTATION
GPEx® Cell Line Engineering Case Studies using Multiple Mammalian Cell Lines 

Greg Bleck, Ph.D., R&D Platform Director, Biologics, Catalent Pharma SolutionsThrough case study examples, attendees will gain an understanding of how GPEx® Cell Line Engineering was incorporated into several cell line expression/ product development projects.  The presentation will discuss the challenges of the overall projects, procedures completed, analysis of data, insights gained, and final conclusions that demonstrate how GPEx® technology was used to generate mammalian cells with high yields and stability, which will help speed the drug to clinic.

 

Innovative Solutions 

2:00 Chairperson’s Remarks

Yvonne Franke, Ph.D., Senior Scientific Manager, Structural Biology, Genentech, Inc.

» KEYNOTE PRESENTATION: 

2:05 Understanding the Secretory Capacity of Pichia pastoris by Single Cell Analysis

Christopher LoveJ. Christopher Love, Ph.D., Associate Professor, Chemical Engineering, MIT

2:35 Maximizing Effectiveness of Protein Expression for Drug Discovery

Yvonne Franke, Ph.D., Senior Scientific Manager, Structural Biology, Genentech, Inc.

We have implemented a high-throughput cloning and protein expression approach which enables us to increase efficiency and effectiveness of successfully producing soluble proteins for drug discovery. Our platform combines strategies such as restriction-independent cloning, plasmid purifications using Phynexus technology, small scale baculovirus infections in multiwell blocks, and analysis of expression levels using Ni-NTA Phytips. This process enables us to quickly assess protein quality and prioritize constructs.

3:05 The SPP (Single Protein Production) System in E. coli; Dual Induction for Creation of Novel Functional Proteins Containing Unnatural Amino Acids

Yojiro Ishida, Researcher, Department of Biochemistry, Robert Wood Johnson Medical School, The Center for Advanced Biotechnology and Medicine

Canavanine (Can), an Arg analogue, is highly toxic and used for some plants to protect themselves. Here, we established an in vivo system to achieve 100% replacement of all Arg residues in a protein with Can in the dual-inducible SPP system with use of His-less MazF and a His auxotroph. Using this system, we successfully created a novel functional protein.

3:35 SRP Pathway Engineering for the Enhanced Production of Antibody in E. coli

Ki Jun Jeong, Ph.D., Professor, Chemical and Biomolecular Engineering, KAIST

In the E. coli host, SRP pathway is not a major pathway and so it has not been widely used for preparative purpose of recombinant proteins including antibody. In this work, we developed efficient SRP pathway by engineering several key components of SRP pathway. With engineered host-vector systems, higher productivity of functional antibody could be achieved in flask cultivation as well as high cell density cultivation.

4:05 End of Conference



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