A label-free immunosensor array using single-chain antibody fragments

Proc Natl Acad Sci U S A. 2005 Oct 11;102(41):14587-92. doi: 10.1073/pnas.0504917102. Epub 2005 Sep 28.

Abstract

We report a microcantilever-based immunosensor operated in static deflection mode with a performance comparable with surface plasmon resonance, using single-chain Fv (scFv) antibody fragments as receptor molecules. As a model system scFv fragments with specificity to two different antigens were applied. We introduced a cysteine residue at the C terminus of each scFv construct to allow covalent attachment to gold-coated sensor interfaces in directed orientation. Application of an array enabled simultaneous deflection measurements of sensing and reference cantilevers. The differential deflection signal revealed specific antigen binding and was proportional to the antigen concentration in solution. Using small, oriented scFv fragments as receptor molecules we increased the sensitivity of microcantilevers to approximately 1 nM.

Publication types

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

MeSH terms

  • Antigens / metabolism*
  • Biosensing Techniques / instrumentation
  • Biosensing Techniques / methods*
  • Cloning, Molecular
  • Enzyme-Linked Immunosorbent Assay
  • Immunoassay / instrumentation
  • Immunoassay / methods*
  • Immunoglobulin Fragments / analysis*
  • Immunoglobulin Fragments / chemistry
  • Immunoglobulin Fragments / isolation & purification
  • Immunoglobulin Fragments / metabolism*

Substances

  • Antigens
  • Immunoglobulin Fragments