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Design of atto-vial based recombinant antibody arrays combined with a planar wave-guide detection system

Author:
  • Sara Ghatnekar-Nilsson
  • Linda Dexlin Mellby
  • Christer Wingren
  • Lars Montelius
  • Carl Borrebaeck
Publishing year: 2007
Language: English
Pages: 540-547
Publication/Series: Proteomics
Volume: 7
Issue: 4
Document type: Journal article
Publisher: John Wiley & Sons

Abstract english

Antibody microarray is a rapidly emerging, powerful approach with great promise within high-throughput proteomics. However, before a truly proteome-wide analysis can be performed, the antibody array format needs to be miniaturized even further in order to enable ultradense arrays to be fabricated. To this end, we have designed and generated proof-of-concept for the first generation of an atto-vial based recombinant antibody array platform. Briefly, we have designed a novel nanostructured substrate using electron beam lithography. Vials, ranging in volume/size from 6 (200 nm in diameter) to 4000 aL (5 mu m in diameter), were fabricated. Human recombinant single-chain Fv antibody fragments, microarray adopted by design, were used as probes. The set-up was interfaced with planar wave-guide technology for evanescant field fluorescence detection. The results showed that protein analytes could be specifically detected in the subzeptomole range for pure systems, using vials down to 57 aL. Further, low-abundant (pg/mL) protein analytes could be detected in directly labeled complex proteomes, such as human whole serum, using 157 aL-vials. Taken together, these results outline the potential of the atto-vial array set-up for miniaturized affinity proteomics-based approaches.

Keywords

  • Condensed Matter Physics
  • Immunology in the medical area
  • microarray
  • antibody
  • atto-vial
  • recombinant

Other

Published
  • ISSN: 1615-9861
Carl Borrebaeck
E-mail: carl [dot] borrebaeck [at] immun [dot] lth [dot] se

Professor

Department of Immunotechnology

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Create Health

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