title: Isolation of scFv antibody fragments against HER2 and CEA tumor antigens from combinatorial antibody libraries derived from cancer patients.
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.
authors.affiliation: Department of Genetics, Faculty of Science, Shahrekord University, P.O. Box 115, Shahrekord, Islamic Republic of Iran; Department of Genetics, Faculty of Science, Tarbiat Modaress University, P.O. Box 14115-175, Tehran, Islamic Republic of Iran; National Institute for Genetic Engineering and Biotechnology, P.O. Box 14155-6343, Tehran, Islamic Republic of Iran.
authors.forename: Hoda
authors.initials: H
authors.lastname: Ayat
authors.forename: Oscar R
authors.initials: OR
authors.lastname: Burrone
authors.forename: Majid
authors.initials: M
authors.lastname: Sadghizadeh
authors.forename: Eissa
authors.initials: E
authors.lastname: Jahanzad
authors.forename: Nasrin
authors.initials: N
authors.lastname: Rastgou
authors.forename: Sarrira
authors.initials: S
authors.lastname: Moghadasi
authors.forename: Mehdi
authors.initials: M
authors.lastname: Arbabi
mesh_terms: D000595:Amino Acid Sequence; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D002272:Carcinoembryonic Antigen / Q000235:genetics / Q000276:immunology*; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D057809:HEK293 Cells; D006801:Humans; D061986:MCF-7 Cells; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000235:genetics / Q000276:immunology*; D019151:Peptide Library; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000276:immunology*; D017386:Sequence Homology, Amino Acid; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000951:Antigens, Neoplasm; D002272:Carcinoembryonic Antigen; D019151:Peptide Library; D018719:Erb-b2 Receptor Tyrosine Kinases; D057127:Single-Chain Antibodies; C508053:ERBB2 protein, human
keywords: Antibody library; CEA; ECD; HER-2/neu; Phage display; carcinoembryonic antigen; control; ctrl; extracellular domain; mAb; monoclonal Antibody; scFv; single chain fragments