The use of wheat-germ lectin-Sepharose for the purification of human haemopexin.
Haemopexin was prepared in 37% yield from normal human serum by a simple procedure involving fractional poly(ethylene glycol) precipitation and subsequent chromatography on DEAE-Sepharose CL-6B. One peak from the ion exchanger consisted of only haemopexin and transferrin. These proteins were separated by chromatography on wheat-germ lectin-Sepharose 6MB. Haemopexin was selectively bound and was subsequently desorbed by N-acetyl-D-glucosamine. No impurities could be detected in the final preparation by immunoelectrophoresis or by immunodiffusion against a range of antisera. The protein gave two partially separated bands in polyacrylamide-gradient-gel electrophoresis, corresponding to apohaemopexin and haem-haemopexin complex.
D000596:Amino Acids / Q000032:analysis; D002846:Chromatography, Affinity / Q000379:methods*; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006466:Hemopexin / Q000032:analysis / Q000302:isolation & purification*; D006801:Humans; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D037102:Lectins; D037121:Plant Lectins; D012685:Sepharose; D014908:Triticum
D000596:Amino Acids; D037102:Lectins; D037121:Plant Lectins; D012685:Sepharose; D006466:Hemopexin