Abstract: A murine monoclonal antibody which selectively binds to the determinant site or specific epitope of glycophorin A exposed on the membrane of the erythrocyte and does not bind to other glycophorins. The monoclonal antibody is coated on a microsphere or substrate of a suitable monodispersed variety and utilized in a separation procedure for recovery of white blood cell subsets without lysing of erythrocytes. The bound microspheres are recovered without adverse depletion of the white blood cell population of the sample. The microsphere or bead-to-cell ratio employed is most acceptable for commercial applications of the invention. The invention thus enables accurate assaying of white blood cell subsets in circulating peripheral blood without resorting to lysing of erythrocytes.
Type:
Grant
Filed:
November 19, 1985
Date of Patent:
June 21, 1988
Assignee:
Coulter Corporation
Inventors:
Kenneth H. Kortright, David E. Hofheinz
Abstract: Detection of an identified human carcinoma tumor antigen in a pathological sample by means of a labelled monoclonal antibody specific to the determinant site on the antigen is enhanced and/or accelerated at an earlier development stage than heretofore achieved by removing a carbohydrate steric hindrance for monoclonal antibody availability to bind the antigen of the tumor for which it is specific. The carbohydrate steric hindrance for monoclonal binding to the antigen is identified as sialic acid. The method of the invention involves selective removal of sialic acid from the antigen's determinant site by enzymatic digestion using neuraminidase.