Fabricating cholyglycine-glucose-6-phosphate dehydrogenase conjugates for cholyglycine detection.
Biotechnol Appl Biochem
; 2020 Jul 07.
Article
em En
| MEDLINE
| ID: mdl-33034915
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, 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.
Texto completo:
1
Bases de dados:
MEDLINE
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Revista:
Biotechnol Appl Biochem
Assunto da revista:
BIOQUIMICA
/
BIOTECNOLOGIA
Ano de publicação:
2020
Tipo de documento:
Article
País de afiliação:
China