Method of producing a cell surface bound lam-1 specific antibody
AssigneeDana-Farber Cancer Institute, Inc.
InventorsThomas F. Tedder, Boris Schleiffenbaum, Olivier Spertini
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Abstract
A shed form of leukocyte adhesion molecule-1 (LAM-1, L-selectin) is present in high levels in human plasma. Quantitative methods of detecting shed LAM-1 (sLAM-1) by Western blot and ELISA analysis are disclosed. Also disclosed are methods for the specific detection of cell-surface bound LAM-1 in the presence of shed LAM-1 and for immunotherapy using monoclonal antibodies reactive with cell-surface bound LAM-1 but not reactive with shed LAM-1. In addition a method of producing an antibody that is reactive with cell-surface bound LAM-1 but not reactive with shed LAM-1 is disclosed.
Claims2
A method for producing an antibody that is reactive with cell-surface bound LAM-1 but not reactive with shed LAM-1, wherein the method comprises the steps of contacting antibody producing cells with a fusion protein comprising the extracellular portion of human leukocyte adhesion molecule-1 (LAM-1) fused to the constant region of a human IgG1 constant region, wherein the fusion protein binds to the anti-LAM1-1 monoclonal antibody produced bv the hybridoma cell line having ATCC accession No. HB 10844, and isolating an antibody that binds to the LAM-1 portion of the fusion protein or to cell surface bound LAM-1, but that does not bind to shed LAM-1.
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