A sensitive enzyme immunoassay for human epidermal growth factor. Determination of hEGF in human serum and urine and pharmacokinetics in mouse.
A sensitive enzyme immunoassay for human epidermal growth factor (hEGF) is described. The anti-hEGF antibody was prepared by immunizing rabbits with hEGF, which was synthesized by Escherichia coli using the genetic engineering technique. The present assay system was based on the sandwiching of an antigen between anti-hEGF F(ab')2 precoated on a 96-well polystyrene plate and beta-D-galactosidase-labeled anti-hEGF Fab'. The range of measurable hEGF by this assay was 0.1-100 pg/well. Recoveries of hEGF added to serum and urine ranged between 94 and 108%. The intra- and inter-assay coefficients of variation were less than 6 and 8%, respectively. The results obtained by this assay method correlated well with those obtained by the radioimmunoassay method. By using this assay, the time course of serum hEGF levels in mice after the various administrations were also examined.
Hayashi
T
T
Institute for Biotechnology Research Laboratories, Wakunaga Pharmaceutical Co., Ltd., Hiroshima, Japan.
Hashimoto
K
K
Sakamoto
S
S
D000818:Animals; D004815:Epidermal Growth Factor / Q000008:administration & dosage / Q000032:analysis* / Q000097:blood / Q000493:pharmacokinetics / Q000652:urine; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D007263:Infusions, Parenteral; D007273:Injections, Intramuscular; D007279:Injections, Subcutaneous; D051379:Mice; D011817:Rabbits; D015203:Reproducibility of Results
D004815:Epidermal Growth Factor