The goal of this proposal is to develop antibody library-based methods which promote efficient identification of clinically relevant tumor specific cell surface antigens, methods which are applicable to the identification of cell type-specific lineage markers in general. The abnormal physiology of tumor cells is reflected in part in the altered chemical and molecular composition of their cell surface. Progressive changes in surface molecule expression allow tumor cells to respond efficiently to external signals for growth and survival, to interact with host tissues, to achieve metastasis, and to avoid immune surveillance. The identification and targeting of tumor-specific cell surface antigens, however, is hampered by the complexity of the epitope space at the tumor cell surface. In addition to proteins, relevant antigens include carbohydrates and other post-translational modification products that cannot be predicted from studies of genomic copy number or mRNA expression levels. This proposal aims to develop combinatiroal antibody library-based strategies that allow efficient identification of tumor specific cell surface antigens for diagnostic, prognostic and therapeutic applications. Specifically, we aim (1) To develop a high throughput subtractive selection strategy based on flow cytometric sorting to significantly improve selection efficiency and to obtain greater numbers of tumor- targeting phage antibodies. (2) To identify clinically relevant tumor antigens by selecting phage antibodies against prostate cancer cells in situ, within their proper stromal contexts. We propose to combine antibody library technology with laser capture microdissection (LCM) to identify antibodies against individual tumor cells on tissue slides. The resulting antibodies will have a very high likelihood of recognizing clinically relevant tumor antigens. (3) To establish the molecular identify of tumor cell surface antigens recognized by tumor-specific antibodies. We have developed a novel combinatorial yeast cDNA display library. Specific antigen-antibody pairs will be identified from this library following flow sorting. In parallel, we will pursue the analysis of tumor antigens by mass spectrometry methods which are capable of analyzing post-translational modifications. This study will increase our knowledge of tumor physiology and will facilitate the design of effective therapy; it will also help determine the exact contribution of post-translational modifications to the final makeup of the tumor epitope space.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA118919-03
Application #
7367198
Study Section
Special Emphasis Panel (ZRG1-RUS-D (50))
Program Officer
Mccarthy, Susan A
Project Start
2006-03-07
Project End
2011-01-31
Budget Start
2008-02-01
Budget End
2009-01-31
Support Year
3
Fiscal Year
2008
Total Cost
$348,376
Indirect Cost
Name
University of California San Francisco
Department
Anesthesiology
Type
Schools of Medicine
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Ha, Kevin D; Bidlingmaier, Scott M; Liu, Bin (2016) Macropinocytosis Exploitation by Cancers and Cancer Therapeutics. Front Physiol 7:381
Sherbenou, Daniel W; Aftab, Blake T; Su, Yang et al. (2016) Antibody-drug conjugate targeting CD46 eliminates multiple myeloma cells. J Clin Invest 126:4640-4653
Bidlingmaier, Scott; Ha, Kevin; Lee, Nam-Kyung et al. (2016) Proteome-wide Identification of Novel Ceramide-binding Proteins by Yeast Surface cDNA Display and Deep Sequencing. Mol Cell Proteomics 15:1232-45
Bidlingmaier, Scott; Liu, Bin (2015) Utilizing Yeast Surface Human Proteome Display Libraries to Identify Small Molecule-Protein Interactions. Methods Mol Biol 1319:203-14
Sherbenou, Daniel W; Behrens, Christopher R; Su, Yang et al. (2015) The development of potential antibody-based therapies for myeloma. Blood Rev 29:81-91
Bidlingmaier, Scott; Liu, Bin (2015) Identification of Posttranslational Modification-Dependent Protein Interactions Using Yeast Surface Displayed Human Proteome Libraries. Methods Mol Biol 1319:193-202
Bidlingmaier, Scott; Liu, Bin (2015) Identification of Novel Protein-Ligand Interactions by Exon Microarray Analysis of Yeast Surface Displayed cDNA Library Selection Outputs. Methods Mol Biol 1319:179-92
Bidlingmaier, Scott; Su, Yang; Liu, Bin (2015) Combining Phage and Yeast Cell Surface Antibody Display to Identify Novel Cell Type-Selective Internalizing Human Monoclonal Antibodies. Methods Mol Biol 1319:51-63
Ha, Kevin D; Bidlingmaier, Scott M; Zhang, Yafeng et al. (2014) High-content analysis of antibody phage-display library selection outputs identifies tumor selective macropinocytosis-dependent rapidly internalizing antibodies. Mol Cell Proteomics 13:3320-31
Behrens, Christopher R; Liu, Bin (2014) Methods for site-specific drug conjugation to antibodies. MAbs 6:46-53

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