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

Ghatnekar-Nilsson, Sara LU ; Dexlin Mellby, Linda LU ; Wingren, Christer LU ; Montelius, Lars LU and Borrebaeck, Carl LU (2007) In Proteomics 7(4). p.540-547
Abstract
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.... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
microarray, antibody, atto-vial, recombinant
in
Proteomics
volume
7
issue
4
pages
540 - 547
publisher
John Wiley & Sons
external identifiers
  • wos:000244807500007
  • scopus:33847683875
ISSN
1615-9861
DOI
10.1002/pmic.200600485
language
English
LU publication?
yes
id
eab74b16-12c2-4983-8165-12ecf9f4afcc (old id 670385)
date added to LUP
2007-12-10 14:41:40
date last changed
2017-01-01 04:41:12
@article{eab74b16-12c2-4983-8165-12ecf9f4afcc,
  abstract     = {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.},
  author       = {Ghatnekar-Nilsson, Sara and Dexlin Mellby, Linda and Wingren, Christer and Montelius, Lars and Borrebaeck, Carl},
  issn         = {1615-9861},
  keyword      = {microarray,antibody,atto-vial,recombinant},
  language     = {eng},
  number       = {4},
  pages        = {540--547},
  publisher    = {John Wiley & Sons},
  series       = {Proteomics},
  title        = {Design of atto-vial based recombinant antibody arrays combined with a planar wave-guide detection system},
  url          = {http://dx.doi.org/10.1002/pmic.200600485},
  volume       = {7},
  year         = {2007},
}