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

Aim To establish antibody sandwich enzyme linked immunoadsorbent assay for determination of recombinant E.coli L asparaginase in rat plasma and study its pharmacokinetics. Methods A 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. Results The 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. Conclusion Antibody 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.