Antibody sandwich enzyme-linked immunoadsorbent assay for determination of recombinant E.coli L-asparaginase in rat plasma and its pharmacokinetics

AimTo establish antibody sandwich enzyme-linked immunoadsorbent assay for determination of recombinant E.coli L-asparaginase in rat plasma and study its pharmacokinetics. MethodsA Japanese white rabbit was immunized with recombinant E.coli L-asparaginase. Immunoglobulin G was separated and purified by using DEAE-cellulose chromatography. Conjugation of horseradish peroxidase to immunoglobulin G was obtained using the two-step glutaraldehyde method. Recombinant E.coli L-asparaginase protein in plasma was measured by antibody sandwich enzyme-linked immunoadsorbent assay. Pharmacokinetic parameters were assessed with model-dependent method. ResultsThe linearities was 1-64 U·L^-1. Concentration-time profile after iv of 1.250, 2.50, 5.00 kU·kg^-1 of recombinant E.coli L-asparaginase fitted with a two-compartment model. The first and terminal elimination T_1/2 were 0.50-0.57 h and 2.45-3.02 h, respectively. The AUC was linearly related to the doses. ConclusionAntibody sandwich enzyme-linked immunoadsorbent assay was constant, reliable, sensitive, and suitable for the determination of recombinant L-asparaginase. Pharmacokinetics of recombinant E.coli L-asparaginase in rats is warranted for the design of future clinical trails.