Fabricating cholyglycine–glucose‐6‐phosphate dehydrogenase conjugates for cholyglycine detection. (11th November 2019)

• Record Type: Journal Article
• Authors: Yang, Mei‐Na; Zhang, Ya‐Fang; Zhi, Gao‐Ying; Gu, Xiao‐Fei; Han, Li; Zhang, Dong‐Hao
• Is Part Of: Biotechnology and applied biochemistry. Volume 67:Number 2(2020)
• Page Start: 257
• Page End: 264
• Publication Date: 2019-11-11
• Subjects: enzyme‐multiplied immunoassay technique -- cholyglycine -- glucose‐6‐phosphate dehydrogenase -- cholyglycine–glucose‐6‐phosphate conjugate; Biotechnology -- Periodicals; Biochemical engineering -- Periodicals; Biochemistry -- Periodicals; Biochemistry -- Periodicals; Genetic Techniques -- Periodicals; Microbiological Techniques -- Periodicals; 660.6
• DOI: 10.1002/bab.1842
• Languages: English
• ISSNs: 0885-4513

Abstract

Abstract: 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. Abstract : CG‐G6PD conjugates for CG detection.