DIRECT QUANTITATION OF PLATELET-ASSOCIATED IGG BY ELECTRO-IMMUNOASSAY

  • 1 January 1982
    • journal article
    • research article
    • Vol. 60  (1) , 54-58
Abstract
An electroimmunoassay was applied to the quantitation of platelet-associated IgG (PAIgG). Protein solubilized by Triton X100 from washed platelets was electrophoresed at pH 5.0 in agarose gels containing carbamoylated rabbit antihuman IgG (pI [isoelectric point] .simeq. 5.0). Because the rabbit antibody is immobilized under these conditions while PAIgG is freely mobile, rocket precipitates were produced, the heights of which were directly proportional to the amount of antigen (PAIgG) present. PAIgG for normal individuals was 4.3 .+-. 1.7 fg[femtogram]/platelet (mean .+-. 2 SD; n = 35). Increased PAIgG levels (direct assay) were found in 27 of 29 patients with a diagnosis of clinically active idiopathic thrombocytopenic purpura (ITP), ranging from 10.5-101.5 fg/platelet. Moderately elevated PAIgG was found in 8 of 10 patients in an early stage of recovery from ITP (range 7.5-9.5 fg/platelet) and in 3 of 6 patients with apparent nonimmune thrombocytopenia (range 14.5-24.0 fg/platelet). Electroimmunoassay for PAIgG can be performed on patients with platelet counts as low as 2000/.mu.l, yields results in < 24 h, is highly reproducible and appears to provide a useful tool for the evaluation of patients with immunologically mediated thrombocytopenia.