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

Electrophoresis. 2016 Jan;37(2):233-8. doi: 10.1002/elps.201500429. Epub 2015 Nov 27.

Abstract

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.

Keywords: Anti-HA; Antibody-Peptide Interaction; CE-FL; YPYDVPDYA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / immunology*
  • Antibodies, Monoclonal / isolation & purification
  • Binding Sites, Antibody
  • Electrophoresis, Capillary / methods*
  • Fluorescence
  • Hybridomas
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Peptides / chemistry
  • Peptides / immunology*
  • Peptides / isolation & purification

Substances

  • Antibodies, Monoclonal
  • Peptides