Enzyme-linked immunosorbent assay (ELISA) for detection of thermostable protease from Pseudomonassp. AFT-36 of dairy origin

A double-antibody sandwich enzyme-linked immunosorbent 'assay was developed using IgG of anti-Pseudomonassp. AFT-36 protease as coating antibody. The assay could detect 0.19 ng/ml of protease in milk and buffer. It could be completed within 5 h. Cross-reactivity studies showed that AFT-36 is immunologically related to six Pseudomonasspecies, while some Pseudomonasspp.produce immunologically unrelated proteases. The results suggest that the assay could be used to detect protease in dairy products.