通过在TSK G3000SW和G3000SWG上进行高速凝胶过滤从纤溶酶处理的人免疫球蛋白中分离Fab和Fc片段。

Isolation of Fab and Fc fragments from a plasmin-treated human immunoglobulin by high-speed gel filtration on TSK G3000SW and G3000SWG.

作者信息

Watabe S, Takada K, Shitori Y, Hashimoto K

出版信息

Vox Sang. 1985;48(1):1-7. doi: 10.1111/j.1423-0410.1985.tb00138.x.

DOI:10.1111/j.1423-0410.1985.tb00138.x

Abstract

A method is described for isolating intact 7S IgG, Fab and Fc fragments from a plasmin-treated immunoglobulin by high-speed gel filtration on a TSK G3000SW or G3000SWG column. The isolated 7S IgG, Fab and Fc fragments reacted with antihuman IgG, Fab and Fc antiserum, respectively, in both Ouchterlony's double immunodiffusion and immunoelectrophoresis. Each of the Fab and Fc fragments formed a single precipitation line, demonstrating their homogeneity. Anticomplementary activities of intact 7S IgG, Fab and Fc fragments were 2.5, less than or equal to 10 and 2.5 mg protein/ml to inhibit 2 units of CH50, respectively, and the diphtheria antitoxin contents were 2.1, 6.0 and 1.5 IU/150 mg protein, respectively. The molecular composition of plasmin-treated immunoglobulin as determined by gel filtration on a TSK G3000SW column was as follows: 7S IgG 38.0 +/- 0.2%; Fab 42.6 +/- 0.1% and Fc fragment 19.3 +/- 0.1%.

摘要

本文描述了一种通过在TSK G3000SW或G3000SWG柱上进行高速凝胶过滤，从经纤溶酶处理的免疫球蛋白中分离完整7S IgG、Fab和Fc片段的方法。在Ouchterlony双向免疫扩散和免疫电泳中，分离出的7S IgG、Fab和Fc片段分别与抗人IgG、Fab和Fc抗血清发生反应。Fab和Fc片段各自形成一条单一沉淀线，表明其均一性。完整7S IgG、Fab和Fc片段的抗补体活性分别为2.5、小于或等于10以及2.5 mg蛋白/ml以抑制2个CH50单位，白喉抗毒素含量分别为2.1、6.0和1.5 IU/150 mg蛋白。通过在TSK G3000SW柱上进行凝胶过滤测定，经纤溶酶处理的免疫球蛋白的分子组成如下：7S IgG 38.0±0.2%；Fab 42.6±0.1%和Fc片段19.3±0.1%。