DNA-Directed Antibody Immobilization for Robust Protein Microarrays: Application to Single Particle Detection 'DNA-Directed Antibody Immobilization

Methods Mol Biol. 2017:1571:187-206. doi: 10.1007/978-1-4939-6848-0_12.

Abstract

Protein microarrays are emerging tools which have become very powerful in multiplexed detection technologies. A variety of proteins can be immobilized on a sensor chip allowing for multiplexed diagnostics. Therefore, various types of analyte in a small volume of sample can be detected simultaneously. Protein immobilization is a crucial step for creating a robust and sensitive protein microarray-based detection system. In order to achieve a successful protein immobilization and preserve the activity of the proteins after immobilization, DNA-directed immobilization is a promising technique. Here, we present the design and the use of DNA-directed immobilized (DDI) antibodies in fabrication of robust protein microarrays. We focus on application of protein microarrays for capturing and detecting nanoparticles such as intact viruses. Experimental results on Single-particle interferometric reflectance imaging sensor (SP-IRIS) are used to validate the advantages of the DDI method.

Keywords: DNA-directed antibody immobilization; Label free detection; Protein microarrays; SP-IRIS; Single particle detection.

MeSH terms

  • Antibodies, Immobilized*
  • Biosensing Techniques / instrumentation
  • Biosensing Techniques / methods*
  • DNA Probes
  • DNA*
  • Image Processing, Computer-Assisted / methods
  • Immunoconjugates
  • Microfluidics / instrumentation
  • Microscopy / methods
  • Molecular Imaging / instrumentation
  • Molecular Imaging / methods*
  • Protein Array Analysis / instrumentation
  • Protein Array Analysis / methods*
  • Statistics as Topic / methods
  • Viruses / immunology

Substances

  • Antibodies, Immobilized
  • DNA Probes
  • Immunoconjugates
  • DNA