Abstract This invention concerns a new monoclonal antibody (mAb) 4C, recognizing a specific antigen, Leu 200, found in human hematopoietic tissues. The monoclonal antibody precipitates a series of glycoproteins (Leu 200) with a molecular weight range of about 190,000 to 230,000 from both T- and B-cell lines. The series of glycoproteins resolves into four discrete glycoprotein bands, the distribution of which varies according to the cell lines. Bands 3 and 4 predominate in a majority of T-cells whereas band 2 predominates in B cells. Thus, a Leu 200 antigen subset distribution is possible with mAb 4C. Bands 1, 2, 3, 4 have apparent molecular weights of 230K, 215K, 205K, and 190K respectively, with differences in their carbohydrate moieties. mAb 4C is a IgG sub two a, kappa immunoglobulin. 4C has potential use in leukemia, hematopoietic cell differentiation and transplantation diagnoses and therapy.
Method for differentiating between human B cells and T cells in a human hematopoietic specimen which comprises (a) contacting the human hematopoietic specimen with monoclonal antibody 4C (ATCC Accession No. HB 8311) under immunoprecipitating conditions so as to allow the formation of 4C: Leu 200 glycoprotein immunoprecipitate complexes: (b) analyzing the immunoprecipitate complexes by SDS-PAGE to determine the presence of Leu 2 glycoproteins having molecular weights of approximately 230 kD, 215 kD, 205 kD and 190 kD; and (c) differentiating B cells from T cells wherein B cells are indicated when the 215 kD glycoprotein predominates in appearance over the 205 kD and 190 kD glycoproteins, are wherein T cells and indicated when the 205 kD and 190 kD glycoproteins predominate over the 215 kD glycoprotein.
Claim 2
Dependent Claim
126 words
Method for differentiating between human B cells and T cells in a human hematopoietic specimen which comprises (a) contacting the human hematopoietic specimen with monoclonal antibody 4C (ATCC Accession No. HB 8311) under immunoprecipitating conditions so as to allow the formation of 4C: Leu 200 glycoprotein immunoprecipitate complexes: (b) analyzing the immunoprecipitate complexes by SDS-PAGE to determine the presence of Leu 2 glycoproteins having molecular weights of approximately 230 kD, 215 kD, 205 kD and 190 kD; and (c) differentiating B cells from T cells wherein B cells are indicated when the 215 kD glycoprotein predominates in appearance over the 205 kD and 190 kD glycoproteins, are wherein T cells and indicated when the 205 kD and 190 kD glycoproteins predominate over the 215 kD glycoprotein.
Claim 3
Dependent Claim
24 words
Method of claim 1 wherein the hematopoietic specimen to be contacted with the monoclonal antibody is an NP-40 extract of -GlcN-labelled human hematopoietic specimen.
Claim 4
Dependent Claim
31 words
Method of claim 1 wherein the Leu 200 glycoproteins are treated with neuraminadase resulting in an increased electrophoretic mobility in the 205 kD and 190 kD glycoproteins relative to untreated material.
Claim 5
Dependent Claim
27 words
Method of claim 1 wherein the 205 kD and 190 kD glycoproteins have a higher sialic acid content relative to the 230 kD and 215 kD glycoproteins.