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Design of recombinant antibody microarrays for serum protein profiling: Targeting of complement proteins

Ingvarsson, Johan LU ; Larsson, Anette; Sjöholm, Anders LU ; Truedsson, Lennart LU ; Jansson, Bo LU ; Borrebaeck, Carl LU and Wingren, Christer LU (2007) In Journal of Proteome Research 6(9). p.3527-3536
Abstract
Antibody-based microarrays is a novel technology with great promise for high-throughput proteomics. The process of designing high-performing arrays has, however, turned out to be challenging. Here, we have designed the next generation of a human recombinant scFv antibody microarray platform for protein expression profiling of nonfractionated biotinylated human plasma and serum proteomes. The setup, based on black polymer Maxisorb slides interfaced with a fluorescent-based read-out system, was found to provide specific, sensitive (subpicomolar (pM) range) and reproducible means for protein profiling. Further, a chip-to-chip normalization protocol critical for comparing data generated on different chips was devised. Finally, the microarray... (More)
Antibody-based microarrays is a novel technology with great promise for high-throughput proteomics. The process of designing high-performing arrays has, however, turned out to be challenging. Here, we have designed the next generation of a human recombinant scFv antibody microarray platform for protein expression profiling of nonfractionated biotinylated human plasma and serum proteomes. The setup, based on black polymer Maxisorb slides interfaced with a fluorescent-based read-out system, was found to provide specific, sensitive (subpicomolar (pM) range) and reproducible means for protein profiling. Further, a chip-to-chip normalization protocol critical for comparing data generated on different chips was devised. Finally, the microarray data were found to correlate well with clinical laboratory data obtained using conventional methods, as demonstrated for a set of medium abundant (micromolar (mu M) to nanomolar (nM) range) protein analytes in serum and plasma samples derived from healthy and complement-deficient individuals. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
profiling, protein expression, recombinant antibody microarrays, serum proteomics, complement proteins
in
Journal of Proteome Research
volume
6
issue
9
pages
3527 - 3536
publisher
The American Chemical Society
external identifiers
  • wos:000249371000020
  • scopus:34948911792
ISSN
1535-3893
DOI
10.1021/pr070204f
project
CREATE Health
language
English
LU publication?
yes
id
41f28b4c-4f5c-4ba5-9d55-146223f3a768 (old id 687896)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=17696517&dopt=Abstract
date added to LUP
2007-12-13 09:01:56
date last changed
2017-07-09 03:37:26
@article{41f28b4c-4f5c-4ba5-9d55-146223f3a768,
  abstract     = {Antibody-based microarrays is a novel technology with great promise for high-throughput proteomics. The process of designing high-performing arrays has, however, turned out to be challenging. Here, we have designed the next generation of a human recombinant scFv antibody microarray platform for protein expression profiling of nonfractionated biotinylated human plasma and serum proteomes. The setup, based on black polymer Maxisorb slides interfaced with a fluorescent-based read-out system, was found to provide specific, sensitive (subpicomolar (pM) range) and reproducible means for protein profiling. Further, a chip-to-chip normalization protocol critical for comparing data generated on different chips was devised. Finally, the microarray data were found to correlate well with clinical laboratory data obtained using conventional methods, as demonstrated for a set of medium abundant (micromolar (mu M) to nanomolar (nM) range) protein analytes in serum and plasma samples derived from healthy and complement-deficient individuals.},
  author       = {Ingvarsson, Johan and Larsson, Anette and Sjöholm, Anders and Truedsson, Lennart and Jansson, Bo and Borrebaeck, Carl and Wingren, Christer},
  issn         = {1535-3893},
  keyword      = {profiling,protein expression,recombinant antibody microarrays,serum proteomics,complement proteins},
  language     = {eng},
  number       = {9},
  pages        = {3527--3536},
  publisher    = {The American Chemical Society},
  series       = {Journal of Proteome Research},
  title        = {Design of recombinant antibody microarrays for serum protein profiling: Targeting of complement proteins},
  url          = {http://dx.doi.org/10.1021/pr070204f},
  volume       = {6},
  year         = {2007},
}