Development of a new parallelized, optical biosensor platform for label-free detection of autoimmunity-related antibodies

Anal Bioanal Chem. 2014 May;406(14):3305-14. doi: 10.1007/s00216-013-7504-y. Epub 2013 Nov 27.

Abstract

Autoimmune diseases are characterized by the presence of autoantibodies in serum of affected patients. The heterogeneity of autoimmune relevant antigens creates a variety of different antibodies, which requires a simultaneous detection mode. For this reason, we developed a tool for parallelized, label-free, optical detection that accomplishes the characterization of multiple antigen-antibody interactions within a single measurement on a timescale of minutes. Using 11-aminoundecyltrimethoxysilane, we were able to immobilize proteinogenic antigens as well as an amino-functionalized cardiolipin on a glass surface. Assay conditions were optimized for serum measurements with a single spot antigen chip on a single spot 1-λ detection system. Minimized background signal allows a differentiation between patients and healthy controls with a good sensitivity and specificity. Applying polarized imaging reflectometric interference spectroscopy, we evaluated samples from three APS patients and three control subjects for this proof-of-principle and already obtained good results for β2-glycoprotein I and cardiolipin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Anticardiolipin / immunology
  • Antigen-Antibody Reactions
  • Antiphospholipid Syndrome / immunology
  • Autoantibodies / immunology*
  • Autoimmunity / immunology*
  • Biosensing Techniques*
  • Cardiolipins / chemistry*
  • Enzyme-Linked Immunosorbent Assay
  • Equipment Design
  • Glass
  • Humans
  • Microscopy, Interference
  • Prothrombin / chemistry
  • Sensitivity and Specificity
  • Silanes / chemistry*
  • Spectrophotometry
  • beta 2-Glycoprotein I / chemistry*

Substances

  • 11-aminoundecyltrimethoxysilane
  • Antibodies, Anticardiolipin
  • Autoantibodies
  • Cardiolipins
  • Silanes
  • beta 2-Glycoprotein I
  • Prothrombin