Purification of complement components by hydrophobic affinity chromatography on phenyl—Sepharose

FEBS Letters

Fontaine, M.¹  Pruvost, L.¹  Al Salihi, A.¹  Ripoche, J.¹ 

[1] Inserm U-78, 543 Chemin de la Bretèque, F-76230 Bois-Guillaume, France

关键词: Complement components C5 and C3;    Purification;    Affinity chromatography;    hydrophobic;    Phenyl—Sepharose;    Native C3;    Separation of C5;    EDTA;    ethylene-diamine-tetraacetic acid;    ϵACA;    ϵ-amino-caproïc acid;    SDS—PAGE;    sodium dodecyl sulfate—polyacrylamide gel electrophoresis;   

DOI  :  10.1016/0014-5793(82)81342-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Human complement components C5 and C3 were purified with 41% and 20% yields, respectively, by euglobulin precipitation, DEAE—Sephacel ion-exchange chromatography and gel filtration. Phenyl—Sepharose chromatography allowed the complete separation of C3 and C5. C3 bound loosely on the resin whereas C5 bound firmly and was eluted with 50% glycerin solution. Gel filtration on Sephacryl S-300 allowed the depletion of C4bp and H that contaminated C5 preparations. Homogeneity of C5 and C3 preparations was demonstrated by SDS—PAGE and immunochemical analysis. C5 and C3 consisted of two chains (α, 110000; β, 75000) linked by disulfide bridges.

【 授权许可】

Unknown