Electrophoretic study of peptide-mediated quantum dot-human immunoglobulin bioconjugation

Electrophoresis. 2013 Sep;34(18):2725-32. doi: 10.1002/elps.201300088.

Abstract

The bioconjugation of quantum dots (QDs) is a key process in their application for bioanalysis as well as imaging. The coupling of QDs with biologically active molecules such as peptides, nucleic acids, and/or antibodies enables their fluorescent labeling, and therefore, selective and sensitive tracking during the bioanalytical process, however, the efficiency of the labeling and preservation of the biological activity of the bioconjugate have to be considered. In this study, a new approach of the bioconjugation of CdTe-QDs and human immunoglobulin employing a small peptide is described. The heptapeptide (HWRGWVC) was synthesized and characterized by mass spectrometry, liquid chromatography, and capillary electrophoresis. Moreover, the peptide was used as a capping agent for QDs synthesis. The CdTe-QDs were synthesized by microwave synthesis (600 W, 20 min) using 3.2 mM CdCl2 and 0.8 mM Na2TeO3. The bioconjugation of QDs capped by this peptide with immunoglobulin was investigated by capillary electrophoresis and magnetic immunoextraction coupled with electrochemical detection by differential pulse voltammetry. Furthermore, the applicability of prepared bioconjugates for fluorescent immunodetection was verified using immobilized goat antihuman IgG antibody.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / blood
  • Antibodies / chemistry*
  • Antibodies / metabolism
  • Antibodies, Immobilized / chemistry
  • Biotechnology / methods*
  • Chickens
  • Electrophoresis, Capillary / methods
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescent Antibody Technique / methods
  • Humans
  • Immunoglobulin G / blood
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / metabolism
  • Immunoglobulins / blood
  • Immunoglobulins / chemistry
  • Immunoglobulins / metabolism
  • Immunomagnetic Separation
  • Models, Molecular
  • Oligopeptides / chemistry*
  • Quantum Dots*

Substances

  • Antibodies
  • Antibodies, Immobilized
  • IgY
  • Immunoglobulin G
  • Immunoglobulins
  • Oligopeptides