Daha M R, Stuffers-Heiman M, Kijlstra A, van Es L A
Immunology. 1979 Jan;36(1):63-70.
The third component of complement from rat plasma was isolated by precipitation with polyethyleneglycol in the presence of benzamidine, followed by chromatography of the precipitate on CM-cellulose, hydroxyapatite, and QAE A50-Sephadex, and gel-filtration on Sephadex G-200 superfine. C3 was isolated in its native form, as assessed by its specific functional activity and its immunoelectrophoretic mobility. The recovery of C3 was between 18 and 26%, and the final material was homogenous on SDS-PAGE analysis. Rat C3 has an apparent molecular weight of 187,000 and it is composed of two non-identical disulphide linked polypeptide chains of 123,000 and 76,000. A monospecific antiserum against rat C3 was produced by immunization of rabbits with purified C3. Zymosan treatment of rat serum resulted in cleavage of C3 and in the appearance of component with a greater mobility during immunoelectrophoresis. The plasma concentration of rat C3 in Wistar rats was 581 +/- 49 microgram/ml (mean +/- 1 S.D.). Interaction of highly purified rat C3 with purified human components B, D, and P resulted in the formation of a C3 convertase suggesting compatibility between human and rat C3.
通过在苯甲脒存在下用聚乙二醇沉淀，随后将沉淀在CM - 纤维素、羟基磷灰石和QAE A50 - 葡聚糖凝胶上进行层析，并在Sephadex G - 200超细凝胶上进行凝胶过滤，从大鼠血浆中分离出补体的第三成分C3。通过其特定功能活性和免疫电泳迁移率评估，C3以其天然形式被分离出来。C3的回收率在18%至26%之间，最终材料在SDS - PAGE分析中是均匀的。大鼠C3的表观分子量为187,000，它由两条通过二硫键连接的不同多肽链组成，分别为123,000和76,000。用纯化的C3免疫兔子产生了针对大鼠C3的单特异性抗血清。用酵母聚糖处理大鼠血清导致C3裂解，并在免疫电泳过程中出现迁移率更高的成分。Wistar大鼠血浆中大鼠C3的浓度为581±49微克/毫升（平均值±1标准差）。高度纯化的大鼠C3与纯化的人成分B、D和P相互作用导致形成C3转化酶，表明人和大鼠C3之间具有兼容性。
