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Molecular & Cellular Proteomics 8:816-826, 2009.
© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Antibody Library-based Tumor Endothelial Cells Surface Proteomic Functional Screen Reveals Migration-stimulating Factor as an Anti-angiogenic Target^*,S

Hai Hu^,, Yuliang Ran^,, Yushan Zhang, Zhuan Zhou, Simon J. Harris^¶, Long Yu, Lixin Sun, Jian Pan, Jun Liu, Jinning Lou^|| and Zhihua Yang^,**

From the State Key Laboratory of Molecular Oncology, Cancer Institute (Hospital), Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100021, China, ^¶ Australian Research Council Special Research Centre for Green Chemistry, Monash University, Clayton, Victoria 3800, Australia, and ^|| Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing 100029, China

Angiogenesis is critical for cancer development and metastasis. Here we have employed a functional antibody library-based proteomic screen to identify proteins that participate in and might be used as therapeutic targets for tumor-related angiogenesis. Mice were immunized with human esophageal cancer endothelial cells (HECEC). The antibody library was established with the mouse spleen cells the serum of which had most anti-angiogenic effect. Monoclonal antibodies were subjected to an immunoreactive and functional screen and monoclonal antibodies that reacted strongly with cell surface antigens of HECECs and influenced their behavior were selected. Antigens that recognized by the antibodies were obtained by immunoprecipitation and then identified by mass spectrometry analysis. Migration-stimulating factor (MSF), the antigen of 1D2 antibody was identified using this approach. Further studies demonstrated that the 1D2 antibody suppressed MSF-effected migration and adhesion of HECECs on fibronectin matrix. Biodistribution assay showed that MSF targeting antibody 1D2 could specifically home to the xenograft with humanized blood vessel. Targeting treatment with 1D2 antibody significantly suppressed tumor growth through inhibition of human tumor-related angiogenesis. These results indicate that the functional antibody library-based proteomic screen can successfully identify proteins that involved in tumor-related angiogenesis and MSF may be a target for the anti-angiogenic treatment of the esophageal cancer.

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