Jump to main content
Jump to site search


Accurate and sensitive detection of dipeptidyl peptidase-IV activity by liquid chromatography with fluorescence detection

Abstract

Dipeptidyl peptidase-IV (DPP-IV), important prolyl-specific peptidases in the serine hydrolase family, hydrolyzes a large number of endogenous oligopeptides. In clinical studies, the detection of DPP-IV, especially the detection of activity, is particularly important. We previously reported a specific fluorescent two-photon probe for DPP-IV. In this study, to further verify that our probe can be used for the DPP-IV activity detection, we developed an accurate and sensitive method for the determination of DPP-IV activity in human tissue microsomes, cell homogenates or cell supernatants by liquid chromatography-fluorescence detection (LC-FD). This method is based on the fluorescence generated by BAN obtained by detecting DPP-IV hydrolysis of GP-BAN. The linearity, sensitivity, recovery, precision and stability of the method were fully validated. The method based on LC-FD has a lower limit of quantitation of BAN (product of DPP-IV) as low as 5 nM (only need 2 μL), which was much lower than the others. The method also demonstrated good accuracy and precision, both variances were less than 15% between intra- and inter-assay. In addition, the method was successfully applied to the determination of DPP-IV activity in various cell homogenates and cell supernatants. The development of the LC-FD method will help to understand the expression and function of DPP-IV in different biological samples in humans in future studies.

Back to tab navigation

Supplementary files

Article information


Submitted
04 Dec 2019
Accepted
09 Jan 2020
First published
09 Jan 2020

Anal. Methods, 2020, Accepted Manuscript
Article type
Paper

Accurate and sensitive detection of dipeptidyl peptidase-IV activity by liquid chromatography with fluorescence detection

H. Ma, X. Qian, J. Zhang, Q. Jin, L. Zou, S. Liu and G. Ge, Anal. Methods, 2020, Accepted Manuscript , DOI: 10.1039/C9AY02610J

Social activity

Search articles by author

Spotlight

Advertisements