RRC ID |
6582
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Author |
Akahori Y, Kurosawa G, Sumitomo M, Morita M, Muramatsu C, Eguchi K, Tanaka M, Suzuki K, Sugiura M, Iba Y, Sugioka A, Kurosawa Y.
|
Title |
Isolation of antigen/antibody complexes through organic solvent (ICOS) method.
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Journal |
Biochem Biophys Res Commun
|
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.
|
Volume |
378(4)
|
Pages |
832-5
|
Published |
2009-1-23
|
DOI |
10.1016/j.bbrc.2008.11.129
|
PII |
S0006-291X(08)02366-8
|
PMID |
19071089
|
MeSH |
Antibodies, Monoclonal / chemistry
Antibodies, Monoclonal / genetics
Antibodies, Monoclonal / isolation & purification*
Antigen-Antibody Complex / chemistry
Antigen-Antibody Complex / genetics
Antigen-Antibody Complex / isolation & purification*
Humans
Membrane Proteins / immunology*
Methods
Peptide Library*
Solvents / chemistry
|
IF |
2.985
|
Times Cited |
18
|
WOS Category
|
BIOPHYSICS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
Resource |
Human and Animal Cells |
|