A solid phase enzyme linked immunofiltration assay for secretory leucocyte proteinase inhibitor.
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.
Melrose
J
J
Raymond Purves Bone and Joint Research Laboratories, University of Sydney at the Royal North Shore Hospital of Sydney, St. Leonards, N.S.W., Australia.
Ghosh
P
P
Jacoby
A
A
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000906:Antibodies; D002358:Cartilage, Articular / Q000737:chemistry / Q000166:cytology; D002478:Cells, Cultured; D003101:Collodion; D005374:Filtration; D006801:Humans; D007124:Immunoenzyme Techniques; D007962:Leukocytes / Q000737:chemistry; D008875:Middle Aged; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D011994:Recombinant Proteins; D053481:Secretory Leukocyte Peptidase Inhibitor; D015842:Serine Proteinase Inhibitors / Q000032:analysis*
D000906:Antibodies; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D011994:Recombinant Proteins; C506814:SLPI protein, human; D053481:Secretory Leukocyte Peptidase Inhibitor; D015842:Serine Proteinase Inhibitors; D003101:Collodion