title: Isolation of antigen/antibody complexes through organic solvent (ICOS) method.
abstract: We developed a method termed ICOS (isolation of antigen-antibody complexes through organic solvent) for comprehensive isolation of monoclonal antibodies (mAbs) bound to molecules on the cell surface. By mixing a large number of phage particles of an antibody (Ab) library with living cells, antigen (Ag)-Ab complexes were formed on the cell surface. The mixture was overlaid on organic solution in a tube and subjected to centrifugation. Phages bound to cells were recovered from the precipitate. The phage fraction isolated turned out to contain mAbs that bind to very heterogeneous epitopes and show strong binding activity to Ags. The ICOS method was applied to isolation of human mAbs that may be therapeutic against cancers. Sixty percent of clones isolated by the screening of a phage Ab library against cancer cells turned out to bind to various kinds of tumor-associated Ags. The precise protocol of ICOS method and the rationale of efficient screening were described.
authors.affiliation: Division of Antibody Project, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi, Japan.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Akahori
authors.forename: Gene
authors.initials: G
authors.lastname: Kurosawa
authors.forename: Mariko
authors.initials: M
authors.lastname: Sumitomo
authors.forename: Miwa
authors.initials: M
authors.lastname: Morita
authors.forename: Chiho
authors.initials: C
authors.lastname: Muramatsu
authors.forename: Keiko
authors.initials: K
authors.lastname: Eguchi
authors.forename: Miho
authors.initials: M
authors.lastname: Tanaka
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Suzuki
authors.forename: Mototaka
authors.initials: M
authors.lastname: Sugiura
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Iba
authors.forename: Atsushi
authors.initials: A
authors.lastname: Sugioka
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Kurosawa
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D000936:Antigen-Antibody Complex / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008565:Membrane Proteins / Q000276:immunology*; D008722:Methods; D019151:Peptide Library; D012997:Solvents / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D008565:Membrane Proteins; D019151:Peptide Library; D012997:Solvents