Abstract
Epidermal growth factor-like domain 7 (EGFL7) has been implicated in promoting solid tumor growth and metastasis via stimulating tumor-associated angiogenesis. The advent of antibody display technology (phage, bacteria, and yeast) led to an enormous revival in the use of antibodies as diagnostic and therapeutic tools for fighting cancer. However, problems with protein folding, posttranslational modification, and codon usage still limit the number of improved antibodies that can be obtained. We describe here the isolation of an EGFL7-specific antibody from a mammalian cell-based full-length antibody display library generated from peripheral blood mononuclear cells of patients with hepatocellular carcinoma. Using a novel vector, contained glycosylphosphatidylinositol anchor and restriction enzyme sites NheI and ClaI, antibody libraries are displayed as whole IgG molecules on the cell surface and screened for specific antigen binding by a combination of magnetic beads and measured by cell ELISA. Anti-EGFL7 antibody was successfully isolated from the library. The mammalian cell-based full-length antibody display library is a great potential application for rapid identification and cloning of human mAbs of targeting hepatocellular carcinoma.
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Acknowledgments
This study was supported by National Key Basic Research Program (973 Program) funded project (2010CB833605), People’s Republic of China. We thank prof. Guo Yajun, International Joint Cancer Institute, The Second Military Medical University, for supplying vector pcDNA3-CH and pcDNA3-CL.
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Li, F., Liu, YH., Li, YW. et al. Human anti-EGFL7 recombinant full-length antibodies selected from a mammalian cell-based antibody display library. Mol Cell Biochem 365, 77–84 (2012). https://doi.org/10.1007/s11010-012-1245-7
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DOI: https://doi.org/10.1007/s11010-012-1245-7