Ehara H, Yoshimoto T, Yamamoto S, Hattori A
Department of Biochemistry, School of Medicine, Tokushima University, Japan.
Biochim Biophys Acta. 1988 May 2;960(1):35-42.
A peroxidase-linked immunoassay of the sandwich type was developed for a quantitative determination of the amount of human cyclooxygenase. Two species of monoclonal antibodies (hPES01 against the human enzyme and PES-5 against the bovine enzyme) were utilized, which recognized different epitopes on the cyclooxygenase of human platelets. The peroxidase activity of the immunoprecipitate was correlated with the amount of cyclooxygenase. The enzyme immunoassay was applied to platelets from 15 normal subjects and a clinical case of platelet cyclooxygenase abnormality with a prolonged bleeding time. Almost the same level of immunoreactive protein was found in platelets of both normal subjects and the patient. However, the solubilized enzyme from the patient's platelets did not transform arachidonic acid to prostaglandin H2 (PGH2) while thromboxane production from PGH2 was observed at a normal level.
开发了一种夹心型过氧化物酶联免疫测定法,用于定量测定人环氧化酶的含量。使用了两种单克隆抗体(针对人酶的hPES01和针对牛酶的PES-5),它们识别人类血小板环氧化酶上的不同表位。免疫沉淀物的过氧化物酶活性与环氧化酶的量相关。该酶免疫测定法应用于15名正常受试者的血小板以及1例血小板环氧化酶异常且出血时间延长的临床病例。在正常受试者和患者的血小板中发现了几乎相同水平的免疫反应性蛋白。然而,患者血小板中溶解的酶不能将花生四烯酸转化为前列腺素H2(PGH2),而从PGH2产生血栓素的水平正常。