一种用于牛C反应蛋白和血清淀粉样蛋白P成分的新纯化方法。

A new purification procedure for bovine C-reactive protein and serum amyloid P component.

Sarikaputi M, Morimatsu M, Syuto B, Saito M, Naiki M

Department of Veterinary Science, National Institute of Health, Tokyo, Japan.

Int J Biochem. 1991;23(10):1137-42. doi: 10.1016/0020-711x(91)90155-g.

Abstract

A new purification procedure was started with salting-out fractionation of serum proteins at 45-75% saturated ammonium sulfate concentration, followed by HE agarose affinity chromatography by which calcium-dependently bound CRP and SAP were purely eluted with EDTA-containing buffer. 2. Pure CRP and SAP were finally separated by DEAE-5PW HPLC. 3. This procedure gave recovery of 15 and 26%, and fold purification of 2650 and 2400 for CRP and SAP, respectively. 4. Each subunit of CRP and SAP had one intrasubunit disulfide bond, determined by reduction and carboxymethylation.

摘要

一种新的纯化方法开始于在45 - 75%饱和硫酸铵浓度下对血清蛋白进行盐析分级分离，随后进行HE琼脂糖亲和层析，通过该层析用含EDTA的缓冲液将钙依赖性结合的CRP和SAP纯洗脱。2. 纯的CRP和SAP最终通过DEAE - 5PW高效液相色谱法分离。3. 该方法对CRP和SAP的回收率分别为15%和26%，纯化倍数分别为2650和2400。4. 通过还原和羧甲基化测定，CRP和SAP的每个亚基都有一个亚基内二硫键。