Breast carcinoma specific antibody selection combining phage display and immunomagnetic cell sorting.
To discover new specific antibodies directed against disseminated carcinoma cells in breast cancer patients, a strategy combining single-chain variable fragment (scFv) phage display and immunomagnetic cell sorting was developed. A selection model, in which ErbB2-expressing breast carcinoma SKBR3 cells are spiked into a 50-fold excess of lymphocytes, was setup. Selection conditions, optimized using the previously characterized ErbB2-specific F5 phage scFv, led to an outstanding phage enrichment yield of 25,000 after only one round. This protocol applied to human nai ve and synthetic phage display antibody libraries led to the selection, in only two rounds, of individual scFv clones (43 out of 46 tested) specific for non-epithelial carcinoma antigens expressed on SKBR3 cells. This strategy is fully applicable to metastatic cells in effusions from breast carcinoma patients and shall lead to the discovery of immunotools crucial for novel diagnostic and therapeutic approaches.
Mazuet
Christine
C
UMR144 CNRS, Research Division, Institut Curie, Paris, France.
Lerouge
Dimitri
D
Poul
Marie-Alix
MA
Blin
Nathalie
N
D000912:Antibodies, Neoplasm / Q000302:isolation & purification*; D001943:Breast Neoplasms / Q000276:immunology*; D005434:Flow Cytometry; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000302:isolation & purification; D018189:Immunomagnetic Separation / Q000379:methods*; D019169:Jurkat Cells; D019151:Peptide Library
D000912:Antibodies, Neoplasm; D007135:Immunoglobulin Variable Region; D019151:Peptide Library