Effect of salt concentration gradient on separation of different types of specific immunoglobulins by ion-exchange chromatography on DEAE cellulose

被引:24
作者
Tishchenko, GA
Bleha, M
Skvor, J
Bostík, T
机构
[1] Acad Sci Czech Republ, Inst Macromol Chem, Prague 16206 6, Czech Republic
[2] Seva Imuno, Karlstejin 26718, Czech Republic
来源
JOURNAL OF CHROMATOGRAPHY B | 1998年 / 706卷 / 01期
关键词
salt concentration gradient; immunoglobulins;
D O I
10.1016/S0378-4347(97)00453-2
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A three-stage process, consisting of an ammonium sulfate precipitation step, dialysis desalination with microporous anion-exchange Neosepta membranes and anion-exchange chromatography on DEAE-cellulose DE-52 was used for the isolation of mouse monoclonal antibodies specific against different antigens. The ascites fluids contained monoclonal antibodies against human IgG, against horseradish peroxidase and against the heavy chain of human IgM. The effect of the salt concentration gradient in the elution buffer was examined with the aim of optimizing chromatographic conditions. The quality of separation of protein zones was determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis under non-reducing conditions. The immunoreactivity of purified monoclonal antibodies was determined by enzyme-linked immunosorbent assay using a solid-phase adsorbed antigens against which each monoclonal antibody type was directed. (C) 1998 Elsevier Science B.V.
引用
收藏
页码:157 / 166
页数:10
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