A solid-phase immunoassay for detection of IgA1 protease activity on agar plates.
An immunoassay was developed which is capable of detecting IgA1 protease positive microorganisms from biologically derived samples grown on agar plates. Activity was detected by overlaying plates with agar containing insolubilized IgA and capturing Fab fragments on nitrocellulose disks coated with anti-lambda light chain. Using cultures of human saliva, a comparison of this assay with a more traditional method involving electrophoresis and Western blotting of individual colony material revealed excellent agreement between the two methods. This assay should be useful in studying the distribution and ecology of organisms producing IgA1 protease, and in assessing its potential as a virulence factor.
Brown
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Department of Oral Biology, University of Florida, Gainesville 32610.
Leak
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D001419:Bacteria / Q000201:enzymology*; D015153:Blotting, Western; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007070:Immunoglobulin A / Q000378:metabolism; D010447:Peptide Hydrolases / Q000032:analysis*; D012463:Saliva / Q000382:microbiology; D012697:Serine Endopeptidases; D013379:Substrate Specificity
D007070:Immunoglobulin A; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; C021433:IgA-specific serine endopeptidase