Serum Leptin Of Monoclonal Antibodies And Its Application To Explore

Three monoclonal antibodies were obtained by the hybridoma technique using recombinanthuman leptin as the antigen, and their characteristics were determined systematically. Further,these antibodies were applied in the detection of leptin concentration in serum.Those thIee McAbs have high specificity, titer, and affinity constant. They all were the tyTeof IgGl. Their titers were between 106--10-7, affinity constant(Ka) 1 .67 X l09L/mol, 3.0 X108L/mol, 7.5 X 108 L/mol, respectively. Antiserum was prepared by immunizing rabbit.Serum-IgG was purified by ion exchange chromatography. It has high titer, l X l0-', and highspecificity. The purified antiserum was linked with HRP with iodine method. The methodologyindicates that it has high quality.A new enzyme-linked immunosorbent assay (ELISA) was deve1oped using monoclonalantibody as the immobilized antibody, and rabbit anti-human leptin serum as the labeledantibody. The minimum detectable concentration by this assay is l6ng/ml. Recovery of leptinadded to serm was betWeen 92.8--106%, CVs within-run and between-run were 8.36%(n=l0)and 10.22%(n=5) respectively.