The fractionation of human plasma proteins. II. The purification of human complement proteins C3, C3u, and C5 by application of affinity chromatography.
We have devised a purification scheme for human complement proteins C3, C3u, and C5. Affinity chromatography is employed to isolate each of these three proteins in a highly purified form. Typical yields were 910, 30, and 53 mg for C3, C3u, and C5, respectively, from 3 liters of human plasma. These yields reflect 31 and 32% recoveries for C3 and C5, respectively. The major advantages of this protocol over other published procedures are the application of affinity chromatography to yield high quality purified complement components, and the fact that the purification of C3, C3u, and C5 is part of a broader scheme designed for the isolation of over 20 plasma proteins from a single batch of plasma.
DiScipio
R G
RG
Department of Immunology, Scripps Research Institute, La Jolla, California 92037.
Sweeney
S P
SP
D001798:Blood Proteins / Q000302:isolation & purification; D005591:Chemical Fractionation; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003176:Complement C3 / Q000302:isolation & purification*; D003182:Complement C5 / Q000302:isolation & purification*; D011415:Complement Factor B / Q000302:isolation & purification; D017242:Complement Factor H / Q000302:isolation & purification; D003165:Complement System Proteins / Q000302:isolation & purification; D016264:Dextran Sulfate; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology
D001798:Blood Proteins; C038245:C3 protein, human; C493495:CFH protein, human; D003176:Complement C3; D003182:Complement C5; D007074:Immunoglobulin G; D017242:Complement Factor H; D003165:Complement System Proteins; D016264:Dextran Sulfate; D011415:Complement Factor B