The antigen-limited nature of microtiter ELISAs requires partial depletion of IgG to permit reliable determination of rabbit serum IgA antibody activity.
The antigen-limiting nature of microtiter ELISAs predicts that antibodies of minor classes may be underestimated when the same specimen contains large amounts of IgG antibodies specific for the same antigen. Such competitive inhibition can be diagnosed from ELISA titration plots. A method is described to eliminate the negative effects of this competition on the detection of IgA antibodies in rabbit serum. The detectability of rabbit serum antibodies to ovalbumin and bovine serum albumin is increased 10-fold by prior treatment of 1:100 dilutions of serum with 1% Cowan I S. aureus. High concns of S. aureus, e.g. 10%, completely deplete serum IgG without loss of IgA. However, concns higher than 1% do not lead to additional improvement in the detectability of IgA antibodies in the systems studied. The method is rapid, inexpensive and shows no non-specific depletion of IgA from either serum or bronchoalveolar lavage fluid.
Butler
J E
JE
Department of Microbiology, University of Iowa Medical School, Iowa City 52242.
Heo
Y
Y
Adams
P
P
Richerson
H B
HB
D000818:Animals; D000942:Antigens, Bacterial / Q000276:immunology; D001667:Binding, Competitive / Q000502:physiology; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007070:Immunoglobulin A / Q000032:analysis*; D007074:Immunoglobulin G / Q000378:metabolism; D010047:Ovalbumin / Q000276:immunology; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000276:immunology; D013211:Staphylococcus aureus / Q000276:immunology
D000942:Antigens, Bacterial; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D012710:Serum Albumin, Bovine; D010047:Ovalbumin