To establish cholyglycine (CG) detection via enzyme‐multiplied immunoassay technique (EMIT), glucose‐6‐phosphate dehydrogenase (G6PD) was used as a reporter enzyme to prepare hapten–enzyme conjugate. Gel electrophoresis and UV scanning demonstrated that G6PD was successfully labeled with cholyglycine, and CG‐G6PD conjugate was obtained. Furthermore, the effects of various parameters on the preparation of CG‐G6PD conjugates were investigated. Consequently, CG amount, nicotinamide adenine dinucleotide, d‐glucose‐6‐phosphate (G6P), phosphate buffer and the pH, and ionic strength of solution had important effects on the residual activity of CG‐G6PD. Moreover, CG amount, the pH, and G6P played important roles in changing CG labeling location on G6PD. Using the CG‐G6PD conjugate as test kit, the cholyglycine–EMIT calibration curve was established, which could be employed in clinical detection of cholyglycine. This study provides some valuable information for preparing hapten–G6PD conjugates.
To establish cholyglycine (CG) detection via enzyme multiplied immunoassay technique (EMIT), glucose‐6‐phosphate dehydrogenase (G6PD) was used as a reporter enzyme to prepare hapten‐enzyme conjugate. Gel electrophoresis and UV scanning demonstrated that G6PD was successfully labeled with cholyglycine and CG‐G6PD conjugate was obtained. Furthermore, the effects of various parameters on the preparation of CG‐G6PD conjugates were investigated. Consequently, CG amount, NADH, D‐glucose‐6‐phosphate (G6P), phosphate buffer and the pH, and ionic strength of solution had important effects on the residual activity of CG‐G6PD. Moreover, CG amount, the pH, and G6P played important roles in changing CG labeling location on G6PD. Using the CG‐G6PD conjugate as test kit, the cholyglycine‐EMIT calibration curve was established, which could be employed in clinical detection of cholyglycine. This study provides some valuable information for preparing hapten‐G6PD conjugates.This article is protected by copyright. All rights reserved
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.