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A solid phase enzyme linked immunofiltration assay for secretory leucocyte proteinase inhibitor
1Raymond Purves Bone and Joint Research Laboratories, University of Sydney at the Royal North Shore Hospital of Sydney, St. Leonards, N.S.W., Australia.
Abstract:
A solid phase enzyme linked immunosorbent filtration assay (ELIFA) has been developed for secretory leucocyte proteinase inhibitor (SLPI) utilising polyclonal anti-recombinant SLPI (anti-rSLPI) and polyclonal anti-bronchial mucus proteinase inhibitor (anti-BLPI) IgG samples. Millipore HATF nitrocellulose 96-well plates were used as receptacles for the assay and a commercial goat anti-rabbit IgG alkaline phosphatase conjugate was used as a secondary antibody for quantitation of levels of primary antibodies bound to rSLPI in the plate wells. Antigen bound to the HATF plates efficiently and the washing/blocking steps were simplified by vacuum filtration of samples resulting in a rapid and convenient assay system. The ELIFA was also sensitive and a detection limit of 0.1 ng SLPI/well was achieved using either anti-SLPI or anti-BLPI as primary antibodies. This assay was used to demonstrate the production of SLPI at moderate levels (0.5-3 ng/ml media) by human articular chondrocytes grown in monolayer culture.