Chromatographic and electrophoretic studies of circulating immune complexes in plasma.
The protein nature of soluble immune complexes from fresh plasma was studied by combining several analytical biochemical techniques. Free immunoglobulins (Ig) G were separated from larger immune complexes by gel permeation chromatography. In a second step, immune complexes, free IgA and IgM were isolated by protein-A and protein-G affinity chromatography and analysed by two-dimensional gel electrophoresis. Sixteen plasma samples from healthy donors were analysed and evaluated visually. Their protein profiles on the gels turned out to be similar, showing only slight quantitative differences. In one case, additional proteins were detected. To prove the ability of the method, immune complexes were analysed from four plasma samples that showed macro creatine kinase type 1, a complex formation between creatine kinase BB and IgG. This methodology can be used for the examination of immune complexes of unknown protein composition in serum or plasma.
Wiederkehr
F
F
Institute of Clinical Chemistry, University Hospital Zürich, Switzerland.
Büeler
M R
MR
Vonderschmitt
D J
DJ
D000328:Adult; D000936:Antigen-Antibody Complex / Q000097:blood*; D009202:Cardiomyopathies / Q000276:immunology; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003402:Creatine Kinase / Q000276:immunology; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D007070:Immunoglobulin A / Q000302:isolation & purification; D007074:Immunoglobulin G / Q000302:isolation & purification; D007075:Immunoglobulin M / Q000302:isolation & purification; D008297:Male; D008875:Middle Aged; D008970:Molecular Weight; D009369:Neoplasms / Q000276:immunology
D000936:Antigen-Antibody Complex; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D003402:Creatine Kinase