In-capillary detection of fast antibody-peptide binding using fluorescence coupled capillary electrophoresis
| Autor: | Ding Shumin, Jianhao Wang, Haifang Qin, Yuqin Qin, Wang Cheli, Pengju Jiang, Chen Yao, Lin Qiu, Li Jinchen, Li Liu, Teng Yiwan |
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| Rok vydání: | 2015 |
| Předmět: |
Capillary action
Clinical Biochemistry Molecular Sequence Data Peptide binding 02 engineering and technology Bioinformatics 01 natural sciences Biochemistry Dissociation (chemistry) Fluorescence Analytical Chemistry Capillary electrophoresis Molar ratio Animals HA-tag Amino Acid Sequence Mice Inbred BALB C Chromatography Hybridomas biology Chemistry 010401 analytical chemistry Antibodies Monoclonal Electrophoresis Capillary 021001 nanoscience & nanotechnology 0104 chemical sciences biology.protein Binding Sites Antibody Antibody 0210 nano-technology Peptides |
| Zdroj: | Electrophoresis. 37(2) |
| ISSN: | 1522-2683 |
| Popis: | Herein, we report a technique for detecting the fast binding of antibody-peptide inside a capillary. Anti-HA was mixed and interacted with FAM-labeled HA tag (FAM-E4 ) inside the capillary. Fluorescence coupled capillary electrophoresis (CE-FL) was employed to measure and record the binding process. The efficiency of the antibody-peptide binding on in-capillary assays was found to be affected by the molar ratio. Furthermore, the stability of anti-HA-FAM-E4 complex was investigated as well. The results indicated that E4 YPYDVPDYA (E4) or TAMRA-E4 YPYDVPDYA (TAMRA-E4) had the same binding priorities with anti-HA. The addition of excess E4 or TAMRA-E4 could lead to partial dissociation of the complex and take a two-step mechanism including dissociation and association. This method can be applied to detect a wide range of biomolecular interactions. |
| Databáze: | OpenAIRE |
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