Biochemical and immunochemical characterization of the antigen-antibody reaction on a non-toxic biomimetic interface immobilized red blood cells of crucian carp and gold nanoparticles

Biosens Bioelectron. 2007 Jan 15;22(6):1116-20. doi: 10.1016/j.bios.2006.04.031. Epub 2006 Jun 19.

Abstract

A special protein assay system based on a highly hydrophilic, non-toxic and conductive biominetic interface has been demonstrated. To fabricate such assay system, red blood cells of crucian carp (RBC) was initially grown on a glassy carbon electrode surface (GCE) deposited nano-sized gold particles (GPs), a second gold nanoparticle layer (NG) was then absorbed on the RBC surface, and finally mammary cancer 15-3 antibody (anti-CA15-3) was attached on the functional RBC surface. A competitive immunoassay format was employed to detect CA15-3 with horseradish peroxidase (HRP)-labeled CA15-3 as tracer and hydrogen peroxide as enzyme substrate. When the immunosensor was incubated into a mixture solution containing HRP-labeled CA15-3 and CA15-3 sample for 1h at 37 degrees C, the amperometric response decreased with the increment of CA15-3 sample concentration. AFM images of the modified layer revealed a uniform distribution of protein and nanogold. In situ QCM and electrochemical measurements demonstrated that the wanted antibody-antigen reactions should occur with high specificity and selectivity. The specific immunoassay system can be developed further to yield sophisticated structures for other proteins.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigen-Antibody Complex / analysis*
  • Antigen-Antibody Complex / immunology
  • Antigen-Antibody Reactions / immunology
  • Biological Assay / methods*
  • Biomimetic Materials / chemistry*
  • Biosensing Techniques / methods*
  • Carps
  • Cells, Cultured
  • Cells, Immobilized
  • Electrochemistry / methods
  • Erythrocytes / immunology*
  • Gold / chemistry
  • Immunoassay / methods*
  • Nanoparticles / chemistry*
  • Nanoparticles / ultrastructure

Substances

  • Antigen-Antibody Complex
  • Gold