Clinical Chemistry
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Clinical Chemistry 29: 151-153, 1983;
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Clinical Chemistry, Vol 29, 151-153, Copyright © 1983 by American Association for Clinical Chemistry

Column enzyme immunoassay for secretory immunoglobulin A in serum

R Yamamoto, S Kimura, S Hattori, Y Ishiguro and K Kato

This enzyme immunoassay for specific measurement of secretory immunoglobulin A concentrations in human serum involves use of a small chromatographic column as a solid-phase. Serum samples are incubated for 2 h with beta-D-galactosidase-labeled antibody to secretory component, then passed through a 0.1-mL Sepharose 4B column containing antibodies to human immunoglobulin A. After the column is washed to remove the unbound label, the buffer in the column is replaced by a solution of o-nitrophenyl-beta-D-galactoside (a beta-D-galactosidase substrate) and incubated at 25 degrees C overnight. The enzyme reaction is stopped by washing the column with sodium carbonate solution, and the absorbance of the eluate is measured at 420 nm. The concentration of secretory immunoglobulin A can be determined with a minimum detectable sensitivity of 3 mg/L, without interference from free immunoglobulin A and secretory component in the same samples.




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Copyright © 1983 by the American Association for Clinical Chemistry.