Purification of anticardiolipin and lupus anticoagulant activities by using cardiolipin immobilized on agarose beads.
We describe a novel method for the purification of aPL, in which pure CL is immobilized on octyl-sepharose beads by hydrophobic interaction. No lipid contamination was present in eluates, and the system could be reutilized three times without loosing extracting capacity. Four patients with antiphospholipid syndrome were studied. A marked decrease in aCL and LA activities was found in all patient plasmas after the passage through a CL-octyl-sepharose column. Both activities were recovered in eluates which contained beta 2-GP-I and IgGs. beta 2-GP-I was also present in normal plasma eluates, which showed no aCL and slight LA activity. This method represents an improvement in the purification of aPL, and could be useful in explaining the mechanism of action of antibodies that are obtained using pure phospholipid as extracting matrix.
Pengo
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V
Dept. of Cardiology, Padua University School of Medicine, Italy.
Biasiolo
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A
D000293:Adolescent; D000328:Adult; D017153:Antibodies, Anticardiolipin / Q000097:blood / Q000302:isolation & purification*; D016736:Antiphospholipid Syndrome / Q000276:immunology; D002308:Cardiolipins; D002847:Chromatography, Agarose; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000302:isolation & purification; D007075:Immunoglobulin M / Q000097:blood / Q000302:isolation & purification; D016682:Lupus Coagulation Inhibitor / Q000097:blood / Q000302:isolation & purification*; D008297:Male; D008875:Middle Aged; D012685:Sepharose / Q000031:analogs & derivatives
D017153:Antibodies, Anticardiolipin; D002308:Cardiolipins; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D016682:Lupus Coagulation Inhibitor; C027176:octyl-sepharose CL-4B; D012685:Sepharose