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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Kim, Young-Myeong Jung, Se-Hui Hong, Seok-Ho Han, Eun-Taek Ha, Kwon-Soo Ji, Su-Hyun Park, Won Sun |
| Description | Author Affiliation: Jung SH ( Department of Molecular and Cellular Biochemistry, Kangwon National University School of Medicine, Chuncheon, Kangwon-Do 24341, Republic of Korea.); Ji SH ( Department of Molecular and Cellular Biochemistry, Kangwon National University School of Medicine, Chuncheon, Kangwon-Do 24341, Republic of Korea.); Han ET ( Department of Medical Environmental Biology and Tropical Medicine, Kangwon National University School of Medicine, Chuncheon, Kangwon-Do 24341, Republic of Korea.); Park WS ( Department of Physiology, Kangwon National University School of Medicine, Chuncheon, Kangwon-Do 24341, Republic of Korea.); Hong SH ( Department of Internal Medicine, Kangwon National University School of Medicine, Chuncheon, Kangwon-Do 24341, Republic of Korea.); Kim YM ( Department of Molecular and Cellular Biochemistry, Kangwon National University School of Medicine, Chuncheon, Kangwon-Do 24341, Republic of Korea.); Ha KS ( Department of Molecular and Cellular Biochemistry, Kangwon National University School of Medicine, Chuncheon, Kangwon-Do 24341, Republic of Korea. Electronic address: ksha@kangwon.ac.kr.) |
| Abstract | Glucose-6-phosphate dehydrogenase (G6PD) regulates nicotinamide adenine dinucleotide phosphate (NADPH) levels and is related to the pathogenesis of various diseases, including G6PD deficiency, type 2 diabetes, aldosterone-induced endothelial dysfunction, and cancer. Therefore, a highly sensitive array-based assay for determining quantitative G6PD activity is required. Here, we developed an on-chip G6PD activity assay using liquid droplet fluorescence arrays. Quantitative G6PD activity was determined by calculating reduced resorufin concentrations in liquid droplets. The limit of detection (LOD) of this assay was 0.162 mU/ml (2.89 pM), which is much more sensitive than previous assays. We used our activity assay to determine kinetic parameters, including Michaelis-Menten constants (Km) and maximum rates of enzymatic reaction (Vmax) for NADP(+) and G6P, and half-maximal inhibitory concentrations (IC50). We successfully applied this new assay to determine G6PD activity in human plasma from normal healthy individuals (n=30) and patients with inflammation (n=30). The inflammatory group showed much higher G6PD activities than did the normal group (p<0.001), with a high area under the curve value of 0.939. Therefore, this new activity assay has the potential to be used for diagnosis of G6PD-associated diseases and utilizing kinetic studies. |
| ISSN | 09565663 |
| Volume Number | 79 |
| e-ISSN | 18734235 |
| Journal | Biosensors and Bioelectronics |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2016-05-15 |
| Publisher Place | Great Britain (UK) |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Enzyme Assays Instrumentation Glucosephosphate Dehydrogenase Blood Lab-on-a-chip Devices Adult Aged Aged, 80 And Over Biosensing Techniques Equipment Design Female Glucose-6-phosphate Metabolism Humans Kinetics Male Middle Aged Nadp Oxazines Chemistry Oxidation-reduction Journal Article Research Support, Non-u.s. Gov't Discipline Biotechnology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Nanoscience and Nanotechnology Medicine Biophysics Biomedical Engineering Biotechnology Electrochemistry |
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