In-capillary detection of fast antibody-peptide binding using fluorescence coupled capillary electrophoresis.

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.