Determination of apolipoprotein A and its constitutive A-I and A-II polypeptides by separate electroimmunoassays.
Determination of apolipoprotein A and its constitutive A-I and A-II polypeptides by separate electroimmunoassays.
复制标题
通过单独的电免疫测定法测定载脂蛋白 A 及其组成型 A-I 和 A-II 多肽。
作者：
M. D. Curry;P. Alaupovic;C. Suenram
电免疫测定法（“火箭”电泳）描述了人血清载脂蛋白A及其组成型A-I和A-II多肽。纯化的脂蛋白A，A-I和A-II用于制备单特异性抗血清和标准化测定。这些特异性、快速（5-8 h）、精密（试验内和试验间变异系数分别为5%和7%）和准确（通过重量法）的试验适用于测定全血清和各种密度等级脂蛋白中的这些多肽。用完整的和脱脂的脂蛋白获得了相当的结果。结果与放射免疫扩散法或放射免疫法所得结果具有良好的相关性。然而，本方法比前者更快速，比后者免疫测定更简单。正常男性和女性血清中A-I和A-II的浓度相似（A-I分别为143 +/- 24和146 +/- 78 mg/dl，A-II分别为78 +/- 17和83 +/- 25 mg/dl）。患有Ila、llb和IV型高脂蛋白血症的受试者的两种多肽浓度相似，而患有I型疾病、卵磷脂：胆固醇酰基转移酶缺乏症和LP-A缺乏症的患者的A-I（0.3-30 mg/dl）和A-II（11-20 mg/dl）浓度最低。血清和高密度脂蛋白中A-I/A-II的摩尔比接近1。
Electroimmunoassays ("rocket" electrophoresis) are described for human serum apolipoprotein A and its constitutive A-I and A-II polypeptides. Purified lipoprotein A, A-I, and A-II were used to prepare monospecific antisera and to standardize assays. These specific, rapid (5-8 h), precise (the within-and between-assay coefficients of variations are 5 and 7%, respectively), and accurate (by gravimetry) assays are applicable to measurement of these polypeptides in whole serum and in various density classes of lipoproteins. Comparable results are obtained with intact and delipidized lipoproteins. Results correlated well with those obtained by radial immunodiffusion or radioimmunoassay. However, the present procedure is more rapid than the former and simpler than the latter immunoassay. Concentrations of A-I and A-II in the serum of normal men and women were similar (143 +/- 24 and 146 +/- 78 mg/dl, respectively, for A-I and 78 +/- 17 and 83 +/- 25 mg/dl for A-II). Subjects with type lla, llb, and IV hyperlipoproteinemias had similar concentrations of both polypeptides, while patients with type I disease, lecithin:cholesterol acyltransferase deficiency and LP-A deficiency had lowest concentrations of A-I (0.3-30 mg/dl) and A-II (11-20 mg/dl). The molar ratio of A-I/A-II in the serum and high-density lipoproteins was close to unity.