Isolation of antigen/antibody complexes through organic solvent (ICOS) method.
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.
Akahori
Yasushi
Y
Division of Antibody Project, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi, Japan.
Kurosawa
Gene
G
Sumitomo
Mariko
M
Morita
Miwa
M
Muramatsu
Chiho
C
Eguchi
Keiko
K
Tanaka
Miho
M
Suzuki
Kazuhiro
K
Sugiura
Mototaka
M
Iba
Yoshitaka
Y
Sugioka
Atsushi
A
Kurosawa
Yoshikazu
Y
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
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D008565:Membrane Proteins; D019151:Peptide Library; D012997:Solvents