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Generation and analyses of human synthetic antibody libraries and their application for protein microarrays

Säll, Anna LU ; Walle, Maria LU ; Wingren, Christer LU ; Müller, Susanne; Nyman, Tomas; Vala, Andrea; Ohlin, Mats LU ; Borrebaeck, Carl A K LU and Persson, Helena LU (2016) In Protein Engineering, Design and Selection 29(10). p.427-437
Abstract

Antibody-based proteomics offers distinct advantages in the analysis of complex samples for discovery and validation of biomarkers associated with disease. However, its large-scale implementation requires tools and technologies that allow development of suitable antibody or antibody fragments in a high-throughput manner. To address this we designed and constructed two human synthetic antibody fragment (scFv) libraries denoted HelL-11 and HelL-13. By the use of phage display technology, in total 466 unique scFv antibodies specific for 114 different antigens were generated. The specificities of these antibodies were analyzed in a variety of immunochemical assays and a subset was further evaluated for functionality in protein microarray... (More)

Antibody-based proteomics offers distinct advantages in the analysis of complex samples for discovery and validation of biomarkers associated with disease. However, its large-scale implementation requires tools and technologies that allow development of suitable antibody or antibody fragments in a high-throughput manner. To address this we designed and constructed two human synthetic antibody fragment (scFv) libraries denoted HelL-11 and HelL-13. By the use of phage display technology, in total 466 unique scFv antibodies specific for 114 different antigens were generated. The specificities of these antibodies were analyzed in a variety of immunochemical assays and a subset was further evaluated for functionality in protein microarray applications. This high-throughput approach demonstrates the ability to rapidly generate a wealth of reagents not only for proteome research, but potentially also for diagnostics and therapeutics. In addition, this work provides a great example on how a synthetic approach can be used to optimize library designs. By having precise control of the diversity introduced into the antigen-binding sites, synthetic libraries offer increased understanding of how different diversity contributes to antibody binding reactivity and stability, thereby providing the key to future library optimization.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
affinity proteomics, phage display technology, protein microarrays, scFv, synthetic antibody libraries
in
Protein Engineering, Design and Selection
volume
29
issue
10
pages
11 pages
publisher
Oxford University Press
external identifiers
  • scopus:84990890044
  • wos:000386205100005
ISSN
1741-0126
DOI
10.1093/protein/gzw042
language
English
LU publication?
yes
id
b31dd1d9-6a8f-4555-8ebe-d985d831e0b1
date added to LUP
2016-11-02 09:01:39
date last changed
2017-07-09 04:54:25
@article{b31dd1d9-6a8f-4555-8ebe-d985d831e0b1,
  abstract     = {<p>Antibody-based proteomics offers distinct advantages in the analysis of complex samples for discovery and validation of biomarkers associated with disease. However, its large-scale implementation requires tools and technologies that allow development of suitable antibody or antibody fragments in a high-throughput manner. To address this we designed and constructed two human synthetic antibody fragment (scFv) libraries denoted HelL-11 and HelL-13. By the use of phage display technology, in total 466 unique scFv antibodies specific for 114 different antigens were generated. The specificities of these antibodies were analyzed in a variety of immunochemical assays and a subset was further evaluated for functionality in protein microarray applications. This high-throughput approach demonstrates the ability to rapidly generate a wealth of reagents not only for proteome research, but potentially also for diagnostics and therapeutics. In addition, this work provides a great example on how a synthetic approach can be used to optimize library designs. By having precise control of the diversity introduced into the antigen-binding sites, synthetic libraries offer increased understanding of how different diversity contributes to antibody binding reactivity and stability, thereby providing the key to future library optimization.</p>},
  author       = {Säll, Anna and Walle, Maria and Wingren, Christer and Müller, Susanne and Nyman, Tomas and Vala, Andrea and Ohlin, Mats and Borrebaeck, Carl A K and Persson, Helena},
  issn         = {1741-0126},
  keyword      = {affinity proteomics,phage display technology,protein microarrays,scFv,synthetic antibody libraries},
  language     = {eng},
  month        = {10},
  number       = {10},
  pages        = {427--437},
  publisher    = {Oxford University Press},
  series       = {Protein Engineering, Design and Selection},
  title        = {Generation and analyses of human synthetic antibody libraries and their application for protein microarrays},
  url          = {http://dx.doi.org/10.1093/protein/gzw042},
  volume       = {29},
  year         = {2016},
}