Schrader W P, Stacy A R
J Biol Chem. 1979 Dec 10;254(23):11958-63.
A sensitive immunoassay for the adenosine deaminase binding protein (complexing protein) of human kidney has been developed. Impetus for the development of the assay was provided by the observations that (a) antibody to complexing protein does not react with the catalytically active adenosine deaminase monomer, and (b) binding of antibody to complexing protein does not affect the binding or catalytic activity of the enzyme monomer. Preformed immune precipitate prepared from rabbit anti-kidney complexing protein serum and goat anti-rabbit gamma-globulin serum is used to selectively insolubilize complexing protein. Quantitation is accomplished by measuring the intrinsic adenosine deaminating activity or adenosine deaminase binding capacity of the protein held in the immune precipitate. As little as 1 ng of kidney complexing protein can be accurately quantitated with the assay. The assay was used to demonstrate that complexing proteins from liver, lung, spleen, fibroblasts, plasma, and urine react with antibody to kidney complexing protein. The shared capacity to bind adenosine deaminase coupled with their antigenic similarity suggests that the complexing proteins of a number of human tissues and body fluids may be products of the same gene.
已开发出一种用于检测人肾腺苷脱氨酶结合蛋白(复合蛋白)的灵敏免疫测定法。开展该测定法的动因源于以下观察结果:(a)复合蛋白抗体不与具有催化活性的腺苷脱氨酶单体发生反应;(b)抗体与复合蛋白的结合不影响酶单体的结合或催化活性。由兔抗肾复合蛋白血清和山羊抗兔γ球蛋白血清制备的预制免疫沉淀物用于选择性地使复合蛋白不溶。通过测量免疫沉淀物中蛋白质的固有腺苷脱氨活性或腺苷脱氨酶结合能力来进行定量。该测定法能够准确测定低至1 ng的肾复合蛋白。该测定法用于证明来自肝脏、肺、脾脏、成纤维细胞、血浆和尿液的复合蛋白与肾复合蛋白抗体发生反应。结合腺苷脱氨酶的共同能力及其抗原相似性表明,许多人体组织和体液中的复合蛋白可能是同一基因的产物。