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Focused antigen capture immunoassay
Author(s) -
Suen Kitty K. Y.,
Lewis W. H. P.
Publication year - 1994
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.11501501169
Subject(s) - nitrocellulose , isoelectric focusing , immunoassay , densitometry , chromatography , coefficient of variation , antigen , antibody , chemistry , microbiology and biotechnology , membrane , biology , immunology , biochemistry , medicine , enzyme
A new approach is described to immunoblotting in which antigens after isoelectric focusing are captured on nitrocellulose membranes coated with a specific antibody. The method has been applied to the analysis of human IgA ϰ and λ from whole blood and serum without prior purification. The visualized bands were quantified by laser densitometry in four p I 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%.