Moffett R B
Biochim Biophys Acta. 1987 Mar 18;912(1):1-8. doi: 10.1016/0167-4838(87)90240-8.
A method of purifying angiotensinogen (C.A. 11002-13-14) from dog plasma using a monoclonal antibody is presented. More than 98% of applied angiotensinogen was sequestered by the 2-C: G7F7 anti-angiotensinogen monoclonal antibody in solid-phase adsorption assays. Employing immunoaffinity column chromatography, sufficient angiotensinogen was purified for subsequent investigation of angiotensinogen molecular heterogeneity. SDS-polyacrylamide gel electrophoretic analysis resolved two molecular weight forms, an alpha-form at Mr 59,000 and a beta-form at Mr 57,000. These alpha- and beta-plasma angiotensinogen forms were separated by anion-exchange HPLC; an additional gamma-form was also identified. Homogeneous dog angiotensinogen, recovered by reverse-phase HPLC, was partially sequenced. The dog angiotensinogen N-terminal sequence was shown to be more similar to rat than to human angiotensinogen. Since neuraminidase reduced the molecular weight of alpha-angiotensinogen by 4000 and beta-angiotensinogen by 2000, these two forms of the prohormone in dog plasma contain two and one carbohydrate side chain, respectively.
本文介绍了一种使用单克隆抗体从犬血浆中纯化血管紧张素原（C.A. 11002 - 13 - 14）的方法。在固相吸附试验中，2 - C: G7F7抗血管紧张素原单克隆抗体可截留超过98%的加入的血管紧张素原。采用免疫亲和柱色谱法，纯化出足够量的血管紧张素原用于后续血管紧张素原分子异质性的研究。SDS - 聚丙烯酰胺凝胶电泳分析解析出两种分子量形式，一种Mr为59,000的α形式和一种Mr为57,000的β形式。这些α和β血浆血管紧张素原形式通过阴离子交换HPLC分离；还鉴定出了另外一种γ形式。通过反相HPLC回收的纯犬血管紧张素原进行了部分测序。结果表明，犬血管紧张素原的N端序列与大鼠的比与人类的更为相似。由于神经氨酸酶使α - 血管紧张素原的分子量降低了4000，使β - 血管紧张素原的分子量降低了2000，因此犬血浆中这两种激素原形式分别含有两条和一条碳水化合物侧链。
