Generation of miniaturized planar ecombinant antibody arrays using a microcantilever-based printer

Nanotechnology. 2014 Jul 11;25(27):275104. doi: 10.1088/0957-4484/25/27/275104. Epub 2014 Jun 24.

Abstract

Miniaturized (Ø 10 μm), multiplexed (>5-plex), and high-density (>100 000 spots cm(-2)) antibody arrays will play a key role in generating protein expression profiles in health and disease. However, producing such antibody arrays is challenging, and it is the type and range of available spotters which set the stage. This pilot study explored the use of a novel microspotting tool, Bioplume(TM)-consisting of an array of micromachined silicon cantilevers with integrated microfluidic channels-to produce miniaturized, multiplexed, and high-density planar recombinant antibody arrays for protein expression profiling which targets crude, directly labelled serum. The results demonstrated that 16-plex recombinant antibody arrays could be produced-based on miniaturized spot features (78.5 um(2), Ø 10 μm) at a 7-125-times increased spot density (250 000 spots cm(-2)), interfaced with a fluorescent-based read-out. This prototype platform was found to display adequate reproducibility (spot-to-spot) and an assay sensitivity in the pM range. The feasibility of the array platform for serum protein profiling was outlined.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / chemistry*
  • Antibodies / immunology
  • Blood Proteins / analysis*
  • Blood Proteins / immunology
  • Humans
  • Microchemistry / instrumentation
  • Microchemistry / methods*
  • Microfluidics / instrumentation
  • Miniaturization
  • Pilot Projects
  • Protein Array Analysis / instrumentation*
  • Protein Array Analysis / methods*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / immunology

Substances

  • Antibodies
  • Blood Proteins
  • Recombinant Proteins