[Antibody sandwich enzyme-linked immunoadsorbent assay for determination of recombinant E. coli L-asparaginase in rat plasma and its pharmacokinetics].
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.
Chen
Jian-hua
JH
Department of Molecular Biology, China Pharmaceutical University, Nanjing 210009, China.
Wu
Wu-tong
WT
Kazuyuki
Hirano
H
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
D011994:Recombinant Proteins; D001215:Asparaginase