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Wednesday, April 30

8:40 Functional Proteomics for Biomarker and Target Discovery
Joshua LaBaer, M.D., Ph.D, Director, Institute of Proteomics, Harvard Medical School
The Harvard Institute of Proteomics (HIP) has initiated a project to create a sequence-verified collection of full-length cDNAs representing human, human pathogen and model organism coding regions, in recombinational vector systems that allow the immediate in-frame transfer of all coding regions into virtually any protein expression vector. These transfers allow the addition of peptide tags to either or both ends of the proteins. This repository, called the FLEXGene Repository (for Full-Length Expression-ready), is enabling the high-throughput (HT) screening of protein function for the entire set (or any customized subset) of genes using any method of in vitro or in vivo expression. Using HT retroviral methods, these clones have been used to identify proteins capable of driving cell migration, altering the morphogenesis of normal epithelial structures, and affecting drug resistance in cells. A novel form of protein microarray, called nucleic acid programmable protein array (NAPPA), has been developed. This method substitutes the printing of proteins on the array with printing cDNAs encoding the proteins. Thus, the array is a DNA array that can be converted into a protein array by adding cell free protein synthesis machinery. This obviates the need to purify proteins, produces human proteins in a mammalian milieu, and avoids concerns about protein stability on the array because the proteins are made just-in-time for assay.

LOOKING TOWARD SCALE-UP AND THE CLINIC

9:10 Efficient Cell-Mediated Cytotoxicity Using Bispecific Single Domain Antibodies for Cancer Therapy 
Patrick Chames, Ph.D., LISM, CNRS
Antibody dependent cell-mediated cytotoxicity (ADCC) is one of the most important modes of action of therapeutic antibodies. To significantly improve the efficacy of this process, we have designed new nanobody-based bispecific antibodies able to redirect human NK cells toward human cancer cells, leading to an efficient cell lysis.

9:40 Shave Months off Clinical Development Timelines Using Simple and Rapid Selection of High-Performance Stable Mammalian Cells Expressing Recombinant Antibodies 
Mr. Andrew Sandford, Vice President, Selexis Inc. 
Technology based on novel human DNA-based genetic elements are enabling the rapid development and selection high-performance stable mammalian cell lines expressing recombinant antibodies. Requiring only standard laboratory equipment, clonal cell lines based on CHOK1 have been selected within weeks of transfection; this approach has been shown by technology licensees to save over 6 months off cell line development timelines. Within 6 weeks of Transfection, we have obtained titers in shake flasks that exceed 2 gm/liter without any media optimization. 

10:10 Coffee Break in the Exhibit Hall

11:10 Improved Antibody-Drug Conjugates for Cancer Therapy
Paul J. Carter, Ph.D., Vice President, Antibody Technologies, Seattle Genetics, Inc.
Anti-CD70 antibody-drug conjugates (ADCs) containing auristatin drugs have potent antitumor activity in xenografts studies of renal cell cancer. The antitumor potency of the ADCs was enhanced by over 2-fold by modifying the IgG1 delivery vehicle to include the 3 mutations, E233P:L234V:L235A, that impair Fc binding to Fcg receptors. This improved antitumor activity correlates with increases in exposure as shown by pharmacokinetic analysis. The ADCs had similar toxicity profiles in rats. The strategy used here for enhancing the therapeutic index of ADCs is potentially broadly applicable to other antibodies as it is independent of the antigen-binding variable domains.

NOVEL HOSTS AND NEW TECHNIQUES

11:40 Production of Recombinant Antibody Fragments in Bacillus megaterium
Stefan Dübel, Ph.D., Technische Universität Braunschweig, Institut für Biochemie und Biotechnologie
The well established production host Escherichia coli relies on the secretion into the periplasmic space for antibody synthesis. Due to the outer membrane of gram-negative bacteria, only a fraction of this material reaches the medium. Recently, the gram-positive bacterium Bacillus megaterium was shown to efficiently secrete recombinant proteins into the growth medium. Here we evaluated it for the recombinant production of antibody fragments. The lysozyme specific single chain Fv (scFv) fragment D1.3 was successfully produced using B. megaterium. The impact of culture medium composition, gene expression time and culture temperatures on the production of functional scFv protein was systematically analyzed. Per L culture supernatant, more than 400 microg of recombinant His6-tagged antibody fragment were purified by one step affinity chromatography. The material produced by B. megaterium showed an increased specific activity compared to material produced in E. coli. High yields of functional scFv antibody fragments can be produced and secreted into the culture medium by B. megaterium, making this production system a reasonable alternative to E. coli.

1:10 Break

1:30 Chairperson’s Remarks

1:35 Ribosome Display of Fc-1 and Selection of Improved Fc-1 Variants 
George Thom, Ph.D, Senior Research Scientist, Discovery Technology, Cambridge Antibody Technology
Ribosome Display is a cell-free translation based technology for protein selection and evolution from large combinatorial protein libraries. We have now employed those Ribosome Display techniques to effectively explore the hFc-1 sequence space in a high-throughput capacity to identify “Hot-Spot” regions attributing to improved effector functions. By using Ribosome Display we have developed a means by which we can evolve and display dimeric Fc regions. Along with a suite of biochemical and cell-based assays we were able to identify those Fc variants with enhanced antibody dependent cellular cytotoxicity (ADCC).

2:05 In Vivo Imaging Technologies Using Engineered Antibodies
Anna M. Wu, Ph.D., Professor and Vice Chair, Department of Molecular and Medical Pharmacology; Director, Cancer Molecular Imaging Program, UCLA Jonsson Comprehensive Cancer Center; Associate Director, Crump Institute for Molecular Imaging, David Geffen School of Medicine at UCLA
The development of highly targeted therapies (including antibody therapeutics) will benefit from knowledge of target expression in vivo - for biological classification of disease, for selection of patients for therapy, and for monitoring response to targeted therapies. Protein engineering allows rapid conversion of antibodies into fragments optimized for in vivo detection. In this talk, tailoring the pharmacokinetics of engineered antibody fragments for radioactive imaging applications such as immunoPET will be discussed. In addition, antibody fusion proteins and immunoconjugates for bioluminescence/fluorescence imaging will be described. Imaging biomarkers in the form of engineered anti-bodies provide a broad method to assess cell surface phenotype of tumors and tissues in vivo.

3:05 Refreshment Break in the Exhibit Hall

3:50 Problem Solving Break-Out Sessions 

4:50 - 6:00 pm Networking Cocktail Reception in the Exhibit Hall 

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