Accurate and sensitive detection of dipeptidyl peptidase-IV activity by liquid chromatography with fluorescence detection
| Autor: | Shu-Qing Liu, Guang-Bo Ge, Li-Wei Zou, Hong Ma, Qiang Jin, Jing Zhang, Xing-Kai Qian |
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| Rok vydání: | 2020 |
| Předmět: |
Accuracy and precision
Oligopeptide animal structures Chromatography 010405 organic chemistry Chemistry General Chemical Engineering Cell General Engineering Endogeny Serine hydrolase 010402 general chemistry 01 natural sciences Fluorescence 0104 chemical sciences Analytical Chemistry medicine.anatomical_structure medicine Microsome Dipeptidyl peptidase IV activity |
| Zdroj: | Analytical Methods. 12:848-854 |
| ISSN: | 1759-9679 1759-9660 |
| Popis: | Dipeptidyl peptidase-IV (DPP-IV), an important prolyl-specific peptidase in the serine hydrolase family, hydrolyzes a large number of endogenous oligopeptides. In clinical studies, the detection of DPP-IV, especially the detection of its 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 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 from BAN obtained from the hydrolysis of GP-BAN by detecting DPP-IV. 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 for BAN (product of DPP-IV), as low as 5 nM (only needs 2 μL), which is much lower than that of the others. This method also demonstrated good accuracy and precision; variances of both were less than 15% between the 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 for future studies. |
| Databáze: | OpenAIRE |
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