Preparation of a monoclonal antibody, SZ-51, that recognizes an alpha-granule membrane protein (GMP-140) on the surface of activated human platelets.
To identify the changes on the platelet plasma membrane after platelet activation, we prepared monoclonal antibodies specific for activated platelets. Hybridoma cell line SZ-51 was screened for antibodies which bound to thrombin activated platelets but not to resting platelets. 125I-SZ-51, and IgG1, bound 780 molecules/platelet on resting platelets. However, it bound 11,000 molecules/platelet on thrombin activated platelets with high affinity (4.2 nmol/L). The isolated protein using affinity chromatography showed a single band in both periodic acid-Schiff and Coomassie blue staining. Immunoblot analysis revealed that SZ-51 reacted with a 140,000 molecular weight protein which was identified to be alpha-granule membrane protein (GMP-140) in radioimmunoassay. These results demonstrate that the antigen recognized by SZ-51 is GMP-140, which is expressed on the plasma membrane of activated platelets.
Wu
G
G
Jiangsu Institute of Hematology, Suzhou Medical College, People's Republic of China.
Xi
X
X
Li
P
P
Chu
X
X
Ruan
C
C
D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D006801:Humans; D015151:Immunoblotting; D007457:Iodine Radioisotopes; D019007:P-Selectin; D015539:Platelet Activation / Q000187:drug effects / Q000276:immunology*; D010979:Platelet Function Tests; D010980:Platelet Membrane Glycoproteins / Q000276:immunology*; D011863:Radioimmunoassay; D013917:Thrombin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D007457:Iodine Radioisotopes; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins; D013917:Thrombin