Immunosensor incorporating anti-His (C-term) IgG F(ab') fragments attached to gold nanorods for detection of His-tagged proteins in culture medium

Sensors (Basel). 2010;10(6):5409-24. doi: 10.3390/s100605409. Epub 2010 Jun 1.

Abstract

Immunosensors based on gold electrodes (electrochemical) or gold discs (optical) modified with 1,6-hexanedithiol, gold nanorods and Anti-His (C-term) monoclonal antibody F(ab') fragment are described. The antigen detected by the sensing platform is a recombinant histidine-tagged silk proteinase inhibitor (rSPI2-His(6)). Electrochemical impedance spectroscopy (EIS) and surface plasmon resonance (SPR) techniques were used as methods for detection of the antigen. This approach allows to detect the antigen protein in concentration of 10 pg per mL (0.13 pM) of culture medium. The immunosensor shows good reproducibility due to covalent immobilization of F(ab') fragments to gold nanorods layer.

Keywords: F(ab’); His-tag; gold electrodes; nanorods.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques / instrumentation
  • Biosensing Techniques / methods*
  • Culture Media / chemistry*
  • Dielectric Spectroscopy / methods
  • Electrodes
  • Gold / chemistry*
  • Gold / pharmacology
  • Histidine / chemistry
  • Histidine / genetics
  • Histidine / immunology*
  • Immunoassay
  • Immunoglobulin Fab Fragments / chemistry*
  • Immunoglobulin Fab Fragments / pharmacology
  • Insect Proteins / analysis*
  • Insect Proteins / chemistry
  • Insect Proteins / genetics
  • Nanotubes / chemistry
  • Pichia
  • Recombinant Proteins / analysis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Surface Plasmon Resonance / methods

Substances

  • Culture Media
  • Immunoglobulin Fab Fragments
  • Insect Proteins
  • Recombinant Proteins
  • silk proteinase inhibitor 2, Galleria mellonella
  • Histidine
  • Gold