Polyethylene glycol precipitates of serum contain a large proportion of uncomplexed immunoglobulins and C3.
High pressure liquid chromatography (HPLC) was used to fractionate redissolved polyethylene glycol (PEG) precipitates isolated from the sera of normal volunteers and from patients with IgA nephropathy (IgAN) and systemic lupus erythematosus (SLE), 2 diseases characterized by elevated levels of circulating immune complexes. The individual fractions were analyzed by solid phase ELISA for IgA, IgM, C3, IgG, and complexes of IgG-IgA and IgG-C3. Although PEG precipitates were enriched for high molecular weight IgA and IgG (presumably bound within CIC), significant amounts of IgM, unbound IgG and C3 were also present. The quantities of the PEG-precipitable proteins did not correlate with their serum concentrations. IgG-IgA and IgG-C3 complexes were found in all precipitates examined, but the levels of complexes were higher in both patient groups. These results indicate that PEG precipitates a considerable quantity of proteins not bound in immune complexes. There appeared to be greater protein precipitation from sera of the patient groups compared to the amount precipitated from the normal sera. These results suggest that an understanding of the mechanism of PEG precipitation may be important in defining abnormalities in IgAN, SLE and perhaps other diseases characterized by elevated levels of CIC. In addition, the possibility of undetected CIC in PEG precipitable material must be considered.
Crowley-Nowick
P A
PA
Department of Microbiology, University of Alabama at Birmingham, AL, USA.
Campbell
E
E
Schrohenloher
R E
RE
Mestecky
J
J
Mestecky
J
J
Jackson
S
S
D000328:Adult; D000936:Antigen-Antibody Complex / Q000097:blood*; D001323:Autoantibodies / Q000097:blood; D001327:Autoimmune Diseases / Q000097:blood*; D001774:Blood Chemical Analysis / Q000379:methods; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid; D003176:Complement C3 / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005922:Glomerulonephritis, IGA / Q000097:blood*; D006031:Glycosylation; D006801:Humans; D007136:Immunoglobulins / Q000302:isolation & purification*; D008180:Lupus Erythematosus, Systemic / Q000097:blood*; D008297:Male; D008875:Middle Aged; D008970:Molecular Weight; D011092:Polyethylene Glycols / Q000494:pharmacology*; D014461:Ultracentrifugation
D000936:Antigen-Antibody Complex; D001323:Autoantibodies; D003176:Complement C3; D007136:Immunoglobulins; D011092:Polyethylene Glycols