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Biologicals ; 41(6): 345-54, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23845693

ABSTRACT

Tumor cells expressing HER-2/neu and CEA antigens are potentially ideal targets for antibody-targeted therapy. In this study, two large human combinatorial libraries have been generated from the lymph nodes of breast cancer patients that express HER2 and CEA antigens in their tumors. These 'immune' libraries have been constructed in two different formats of scFv, differing in the length of the peptide linker connecting the two variable VH and VL domains. Libraries derived from these patients may contain a larger pool of anti-tumor antigen antibodies and are useful repertoire for isolating scFvs against any tumor markers. The results of this study showed that we were successful in obtaining human scFvs against HER-2/neu and CEA. For HER-2, cell-panning strategy was performed and resulted in two scFv binders that detected the complete HER-2 receptor on the cell membrane and internalized to the cells. Also, preliminary ELISA data showed that several anti-CEA scFv binders were isolated by panning.


Subject(s)
Carcinoembryonic Antigen/immunology , Neoplasms/immunology , Peptide Library , Receptor, ErbB-2/immunology , Single-Chain Antibodies/immunology , Amino Acid Sequence , Antigens, Neoplasm/genetics , Antigens, Neoplasm/immunology , Blotting, Western , Carcinoembryonic Antigen/genetics , Cell Line, Tumor , Cytotoxicity, Immunologic/immunology , Enzyme-Linked Immunosorbent Assay , HEK293 Cells , Humans , MCF-7 Cells , Microscopy, Confocal , Molecular Sequence Data , Neoplasms/genetics , Receptor, ErbB-2/genetics , Sequence Homology, Amino Acid , Single-Chain Antibodies/genetics , Single-Chain Antibodies/isolation & purification
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