Polyclonal and monoclonal antibodies against human tissue inhibitor of metalloproteinases (TIMP) and the design of an enzyme-linked immunosorbent assay to measure TIMP

Abstract

Both polyclonal and monoclonal antibodies have been raised against purified human fibroblast tissue inhibitor of metalloproteinases (TIMP) and characterised. Combinations of antibodies were tested for their suitability in a double antibody sandwich ELISA to measure TIMP. Two combinations were applicable to the immunoassay: (i) a monoclonal capture antibody with a polyclonal detecting antibody; (ii) two monoclonal antibodies. The assay range was (i) 2-50 ng/ml and (ii) 5-50 ng/ml of human TIMP. The levels of TIMP in several human body fluids were measured using assay (ii), and values in ng/ml with standard deviations (n-1) obtained as follows: serum, 299 ± 62; plasma, 109 ± 35; amniotic fluid, 724 ± 62; cerebrospinal fluid, 144 ± 156; saliva, 209 ± 103.