Cell surface antigen detection method

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

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435 2, 436507, 436518, 436528, 436811, G01N 5300, G01N 53564

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048106321

ABSTRACT:
Chronic immune thrombocytopenic purpura is due to platelet destruction by circulating anti-platelet antibody. Although autoantibodies against the platelet glycoprotein IIb/IIIa complex and glycoprotein Ib have been demonstrated using various methods, practical assays for detection of platelet-associated or plasma autoantibodies have not been available. I studied 44 patients with chronic immune thrombocytopenic purpura where platelet-associated and plasma autoantibodies against the glycoprotein IIb/IIIa complex and glycoprotein Ib were measured using a newly developed immunobead assay and a previously reported microtiter well assay. Platelet-associated autoantibody was detected using the immunobead assay in nine of 11 patients (81.8%; seven with anti-GPIIb/IIIa, two with anti-GPIb). Plasma autoantibodies were noted in 28 of 44 patients (63.6%; 19 with anti-GPIIb/IIIa, seven with anti-GPIb and two with both). Positive results were noted in 24 of 44 patients using the immunobead assay and in only 12 of 44 using the microtiter well assay, suggesting that solubilization of the platelets prior to antibody addition, as in the microtiter well assay, alters epitope stability. Of the 20 thrombocytopenic control patients studied, all gave negative results using both assays. I conclude that these assays allow detection of autoantibodies in the majority of patients with chronic immune thrombocytopenic purpura confirming the presence of an autoimmune process. In addition, the two assays may differentiate between epitope types.

REFERENCES:
patent: 4695538 (1987-09-01), Cote et al.
Woods, Jr., et al, "Autoantibodies Against Platelet . . . ", Blood 63(2), 368-375 (Feb. 1984).
Millard et al., Blood, 70:1495-1499 (1987).

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