Purification of C-reactive protein.
A concise method was designed for preparation of C-reactive protein (CRP) from pleural effusion. By addition of L-alpha-lecithin to the pleural effusion in the presence of calcium ions, a flocculence of the CRP-lecithin complex formed. Subsequent treatment of the CRP-lecithin complex with chloroform and sodium citrate buffer enabled extraction of the CRP in the buffer layer. This extracted CRP was further purified by sequential treatment on column chromatography of DEAE cellulose (DE52) and gel filtration using Sephacryl S-300. The isolated protein was proved to be native CRP with a high degree of purity, as determined by electrophoretical and immunological analysis. The yield was 41.8% recovery from the starting material. E1%(280) of the CRP preparation was estimated to be 18.75.
D000818:Animals; D002097:C-Reactive Protein / Q000302:isolation & purification*; D002725:Chloroform; D002848:Chromatography, DEAE-Cellulose / Q000379:methods; D002850:Chromatography, Gel / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D010713:Phosphatidylcholines; D010996:Pleural Effusion / Q000378:metabolism; D011817:Rabbits
D010713:Phosphatidylcholines; D002725:Chloroform; D002097:C-Reactive Protein