Development of validated ultrasensitive enzyme-linked immunosorbent assay method of human epidermal growth factor and in vivo pharmacokinetic study

Nepidermin is a recombinant human epidermal growth factor used for wound healing and is known to be potentially effective against mucosal injury induced by anticancer therapies. This study aimed to develop a sensitive, selective, and validated analysis method for determining nepidermin levels in Sprague-Dawley rat serum and organs and to investigate the compound’s distribution profiles after an intravenous (IV) or subcutaneous (SC) injection.

An enzyme-linked immunosorbent assay (ELISA) was used for the sensitive analysis method development, and the method was validated according to the general recommendations for immunoassays. For the pharmacokinetic and organ distribution study, various concentrations of nepidermin were administered to the rats via the IV or SC route. The amounts of nepidermin in the samples were analyzed using the validated analysis method. A highly sensitive ELISA analysis method was developed and proved suitable for the quantification of nepidermin in the blood and organs of rats.

Our study characterized the dose-dependent pharmacokinetics occurring after an IV or SC administration of nepidermin in rats. Nepidermin was mainly distributed to the liver and kidney after the IV or the SC injection, and the proportion of the nepidermin distribution to the gut (compared to the serum) was equal or higher in the SC groups than in the IV groups.

In conclusion, ultra-sensitive analysis method was developed for the quantification of nepidermin in the blood and various organs of rats, and it was successfully applied to precise interpretation of pharmacokinetic results.