title: [Antibody sandwich enzyme-linked immunoadsorbent assay for determination of recombinant E. coli L-asparaginase in rat plasma and its pharmacokinetics].
abstract: OBJECTIVE
  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 i.v. of 1.25, 2.50, 5.00 kU.kg-1 of recombinant E. coli L-asparaginase fitted with a two-compartment model. The first and terminal elimination T1/2 were 0.50-0.57 h and 2.45-3.02 h, respectively. The AUC was linearly related to the doses.
  
  
  CONCLUSIONS
  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.
authors: Department of Molecular Biology, China Pharmaceutical University, Nanjing 210009, China.
authors: Jian-hua
authors: JH
authors: Chen
authors: Wu-tong
authors: WT
authors: Wu
authors: Hirano
authors: H
authors: Kazuyuki
mesh_terms: D000818:Animals; D019540:Area Under Curve; D001215:Asparaginase / Q000097:blood* / Q000493:pharmacokinetics*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000201:enzymology; D008657:Metabolic Clearance Rate; D011817:Rabbits; D011897:Random Allocation; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000097:blood / Q000493:pharmacokinetics
chemical_list: D011994:Recombinant Proteins; D001215:Asparaginase