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2006 Scientific Report

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Van Andel Research

Van Andel Research Institute | Scientific Report Brian Cao, M.D. Laboratory of Antibody Technology 10 Dr. Cao obtained his M.D. from Beijing Medical University, People’s Republic of China, in 1986. On receiving a CDC fellowship award, he was a visiting scientist at the National Center for Infectious Diseases, Centers for Disease Control and Prevention (1991–1994). He next served as a postdoctoral fellow at Harvard (1994–1995) and at Yale (1995–1996). From 1996 to 1999, Dr. Cao was a Scientist Associate in charge of the Monoclonal Antibody Production Laboratory at the Advanced BioScience Laboratories–Basic Research Program at the National Cancer Institute–Frederick Cancer Research and Development Center, Maryland. Dr. Cao joined VARI as a Special Program Investigator in June 1999. Staff Laboratory Staff Ping Zhao, M.S. Tessa Grabinski, B.S. Students Students Qin Hao Xin Wang Aixia Zhang Jin Zhu Visiting Scientists

VARI | 2006 Research Interests Antibodies are primary tools in several areas of biomedical sciences, including basic research, diagnostics, and molecular therapeutics. In basic biomedical research, the characterization and analysis of almost any molecule involves the production of specific monoclonal or polyclonal antibodies that react with it. The production and sale of antibodies for research purposes is a multibillion-dollar industry worldwide. Antibodies are also widely used in diagnostic applications for clinical medicine. ELISA and radioimmunoassay systems are antibody-based. Analysis of cells and tissues in pathology laboratories includes the use of antibodies on tissue sections and in flow cytometry analyses. Overall, antibody diagnostics also form a multibillion-dollar industry. Antibodies are making rapid inroads into medical therapeutics as well. The success of anti–tumor necrosis factor in the treatment of rheumatoid arthritis and inflammatory bowel disease; anti–B cell antibodies in the treatment of lymphoma; and antibodies against oncogene products in the treatment of breast cancer are only the first examples of what is emerging as a broad new class of therapeutic agents. The monoclonal antibody market is one of the fastest growing, and it has the potential to reach .3 billion by 2010, driven by technological evolution from chimeric and humanized to fully human antibodies. To speed up antibody-related translational research at VARI, the Antibody Technology laboratory has developed several technologies over the last few years: 1) state-of-the-art monoclonal antibody (mAb) production and characterization, followed by scaled-up production and purification; 2) antibody-binding-site epitope mapping using a phage-display peptide library; 3) construction of a human-antibody-fragment phage-display library and screening of specific fragments from the library; and 4) murine-human chimeric antibody generation and murine antibody humanization. 11 Functioning as an antibody production core facility, this lab has extensive capabilities. The technologies and services available in the core include antigen preparation and animal immunization; peptide design and coupling to protein carriers; consultation on protein expression and purification; DNA immunization (gene-gun technology); immunization with living or fixed cells; conventional antigen/ adjuvant preparation; immunization of a wide range of antibody-producing models (including mice, rats, rabbits, human cells, and transgenic or knock-out mice); and in vitro immunization. Our work also includes the generation of hybridomas from spleen cells of immunized mice, rats, and rabbits; hybridoma expansion and subcloning; cryopreservation of hybridomas secreting mAbs; isotyping of mAbs; ELISA screening of hybridoma supernatants; mAb characterization by immunoprecipitation, Western blot, immunohistochemistry, immunofluorescence staining, FACS, and in vitro bioassays; production of bulk quantities of mAbs using high-density cell culture; purification of mAbs on FPLC affinity columns; generation of bi-specific mAbs by secondary fusion; mAb affinity column preparation to purify antigen; conjugation of mAbs to enzymes, biotin/streptavidin, or fluorescent reporters; and development of detection methods/kits such as sandwich ELISA. Over 100 hybridomas against more than 10 unique proteins have been generated and characterized in the past year. In addition, the Antibody Technology lab has established contract services to local biotechnology companies to generate, characterize, produce, and purify mAbs for their research and for diagnostic kit development.

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