Immunoaffinity chromatographic purification of Russell's viper venom factor X activator using elution in high concentrations of magnesium chloride.
RVV-X, the factor X activator from Russell's viper venom, has been isolated using affinity chromatography on agarose columns of a monoclonal antibody specific for this enzyme. Upon testing acid, alkaline, and high concentrations of MgCl2 for elution, it was found that use of high concentrations of MgCl2 was most effective in elution of RVV-X. It was nondenaturing and yielded 90% recovery of homogeneous enzyme without measurable contamination by other proteins of the venom.
Durkee
K H
KH
Department of Biochemistry, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, North Carolina 27157-1016.
Roh
B H
BH
Doellgast
G J
GJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000915:Antibody Affinity; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000276:immunology / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007163:Immunosorbent Techniques; D015636:Magnesium Chloride; D008666:Metalloendopeptidases; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008970:Molecular Weight; D014757:Viper Venoms / Q000201:enzymology*
D000911:Antibodies, Monoclonal; D014757:Viper Venoms; D015636:Magnesium Chloride; D010450:Endopeptidases; D008666:Metalloendopeptidases; C033858:russellysin