Focused antigen capture immunoassay.
A new approach is described to immunoblotting in which antigens after isoelectric focusing are capture on nitrocellulose membranes coated with a specific antibody. The method has been applied to the analysis of human IgA kappa and lambda from whole blood and serum without prior purification. The visualized bands were quantified by laser densitometry in four pI ranges. It is concluded that, under the conditions used, close to 100% of serum IgA is captured. The method has a coefficient of variation of 4.5-12.8%, mean 8.0%.
Department of Health Sciences, Hong Kong Polytechnic, Hung Hom, Kowloon.
K K
KK
Suen
W H
WH
Lewis
D000937:Antigen-Antibody Reactions; D000941:Antigens; D001800:Blood Specimen Collection / Q000379:methods; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting / Q000379:methods; D007070:Immunoglobulin A / Q000097:blood* / Q000302:isolation & purification; D007145:Immunoglobulin kappa-Chains / Q000302:isolation & purification*; D007146:Immunoglobulin lambda-Chains / Q000302:isolation & purification*; D007202:Indicators and Reagents; D007525:Isoelectric Focusing / Q000379:methods
D000941:Antigens; D007070:Immunoglobulin A; D007145:Immunoglobulin kappa-Chains; D007146:Immunoglobulin lambda-Chains; D007202:Indicators and Reagents