TY - JOUR
T1 - Quantitation and characterization of human platelet glycoprotein IIIa by radioimmunoassay
AU - Cierniewski, Czeslaw S.
AU - Niewiarowski, Stefan
AU - Hershock, Diane
AU - Rucinski, Boguslaw
AU - Schmaier, Alvin H.
N1 - Funding Information:
The skillfull technical assistance of Ms. Anette Eckardt is gratefully acknowledged. We wish to thank Dr. Si-Yin Chung for the preparation of the 66 kDa protein. The study was supported in part by NIH grants HL15226, HL14217, HL19055 and HL00694, and by grant 1538 from the Council for Tobacco Research.
PY - 1987/4/16
Y1 - 1987/4/16
N2 - Glycoprotein IIIa was quantitated in human platelets by radioimmunoassay using antisera specific to platelet membranes and purified glycoprotein IIIa. Glycoprotein IIIa and glycoprotein IIb were isolated from washed platelets by Triton X-114 extraction followed by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Radioiodinated glycoprotein IIIa was further purified by affinity chromatography on Lentil lectin-Sepharose 4B. Purified glycoprotein IIb showed little crossreactivity with 125I-labeled glycoprotein IIIa using the anti-platelet membrane or anti-glycoprotein IIIa antisera on a competition inhibition radioimmunoassay. The expression of glycoprotein IIIa epitopes were the same for the purified glycoprotein IIIa and glycoprotein IIIa in Triton X-100 solubilized platelets. A 66 kDa protein derived from glycoprotein IIIa by limited proteolysis of platelet membranes also expressed the same epitopes as intact glycoprotein IIIa. Solubilized platelets contained approximately 16 μg of total glycoprotein IIIa antigen per 109 cells. The level of glycoprotein IIIa determined by radioimmunoassay in one patient with Glanzmann's thrombasthenia amounted to 6.7% of normal and it was close to the values obtained by other methods.
AB - Glycoprotein IIIa was quantitated in human platelets by radioimmunoassay using antisera specific to platelet membranes and purified glycoprotein IIIa. Glycoprotein IIIa and glycoprotein IIb were isolated from washed platelets by Triton X-114 extraction followed by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Radioiodinated glycoprotein IIIa was further purified by affinity chromatography on Lentil lectin-Sepharose 4B. Purified glycoprotein IIb showed little crossreactivity with 125I-labeled glycoprotein IIIa using the anti-platelet membrane or anti-glycoprotein IIIa antisera on a competition inhibition radioimmunoassay. The expression of glycoprotein IIIa epitopes were the same for the purified glycoprotein IIIa and glycoprotein IIIa in Triton X-100 solubilized platelets. A 66 kDa protein derived from glycoprotein IIIa by limited proteolysis of platelet membranes also expressed the same epitopes as intact glycoprotein IIIa. Solubilized platelets contained approximately 16 μg of total glycoprotein IIIa antigen per 109 cells. The level of glycoprotein IIIa determined by radioimmunoassay in one patient with Glanzmann's thrombasthenia amounted to 6.7% of normal and it was close to the values obtained by other methods.
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U2 - 10.1016/0304-4165(87)90090-0
DO - 10.1016/0304-4165(87)90090-0
M3 - Article
C2 - 3828395
AN - SCOPUS:0023140643
SN - 0304-4165
VL - 924
SP - 216
EP - 224
JO - BBA - General Subjects
JF - BBA - General Subjects
IS - 1
ER -