Breast carcinoma specific antibody selection combining phage display and immunomagnetic cell sorting

• Published in: Biochemical and biophysical research communications 2006-09, Vol.348 (2), p.550-559
• Main Authors: Mazuet, Christine, Lerouge, Dimitri, Poul, Marie-Alix, Blin, Nathalie
• Format: Article
• Language: English
• Subjects: Antibodies, Neoplasm - isolation & purification; Breast carcinoma; Breast Neoplasms - immunology; Flow Cytometry; Humans; Immunoglobulin Variable Region - isolation & purification; Immunomagnetic cell sorting; Immunomagnetic Separation - methods; Jurkat Cells; Peptide Library; Phage display antibody library; Single-chain variable fragment
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2006.07.087

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 naı¨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.