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Advancing the global proteome survey platform by using an oriented single chain antibody fragment immobilization approach.

Säll, Anna LU ; Persson, Helena LU ; Ohlin, Mats LU ; Borrebaeck, Carl LU and Wingren, Christer LU (2016) In New Biotechnology 33(5A). p.503-513
Abstract
Increasing the understanding of a proteome and how its protein composition is affected by for example different diseases, such as cancer, has the potential to improve strategies for early diagnosis and therapeutics. The Global Proteome Survey or GPS is a method that combines mass spectrometry and affinity enrichment with the use of antibodies. The technology enables profiling of complex proteomes in a species independent manner. The sensitivity of GPS, and other methods relying on affinity enrichment, is largely affected by the activity of the exploited affinity reagent. We here present an improvement of the GPS platform by utilizing an antibody immobilization approach which ensures a controlled immobilization process of the antibody to... (More)
Increasing the understanding of a proteome and how its protein composition is affected by for example different diseases, such as cancer, has the potential to improve strategies for early diagnosis and therapeutics. The Global Proteome Survey or GPS is a method that combines mass spectrometry and affinity enrichment with the use of antibodies. The technology enables profiling of complex proteomes in a species independent manner. The sensitivity of GPS, and other methods relying on affinity enrichment, is largely affected by the activity of the exploited affinity reagent. We here present an improvement of the GPS platform by utilizing an antibody immobilization approach which ensures a controlled immobilization process of the antibody to the magnetic bead support. More specifically, we make use of an antibody format that enables site-directed biotinylation and use this in combination with streptavidin coated magnetic beads. The performance of the expanded GPS platform was evaluated by profiling yeast proteome samples. We demonstrate that the oriented antibody immobilization strategy increases the ability of the GPS platform and results in larger fraction of functional antibodies. Additionally, we show that this new antibody format enabled in-solution capture, i.e. immobilization of the antibodies after sample incubation. A workflow has been established that permit the use of an oriented immobilization strategy for the GPS platform. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
New Biotechnology
volume
33
issue
5A
pages
503 - 513
publisher
Elsevier
external identifiers
  • pmid:26703809
  • scopus:84964595372
  • wos:000378026000004
ISSN
1876-4347
DOI
10.1016/j.nbt.2015.12.001
language
English
LU publication?
yes
id
4ac1cb76-a4d4-4d32-b9f3-f6ece481182f (old id 8503583)
date added to LUP
2016-01-08 10:22:30
date last changed
2017-09-18 11:28:20
@article{4ac1cb76-a4d4-4d32-b9f3-f6ece481182f,
  abstract     = {Increasing the understanding of a proteome and how its protein composition is affected by for example different diseases, such as cancer, has the potential to improve strategies for early diagnosis and therapeutics. The Global Proteome Survey or GPS is a method that combines mass spectrometry and affinity enrichment with the use of antibodies. The technology enables profiling of complex proteomes in a species independent manner. The sensitivity of GPS, and other methods relying on affinity enrichment, is largely affected by the activity of the exploited affinity reagent. We here present an improvement of the GPS platform by utilizing an antibody immobilization approach which ensures a controlled immobilization process of the antibody to the magnetic bead support. More specifically, we make use of an antibody format that enables site-directed biotinylation and use this in combination with streptavidin coated magnetic beads. The performance of the expanded GPS platform was evaluated by profiling yeast proteome samples. We demonstrate that the oriented antibody immobilization strategy increases the ability of the GPS platform and results in larger fraction of functional antibodies. Additionally, we show that this new antibody format enabled in-solution capture, i.e. immobilization of the antibodies after sample incubation. A workflow has been established that permit the use of an oriented immobilization strategy for the GPS platform.},
  author       = {Säll, Anna and Persson, Helena and Ohlin, Mats and Borrebaeck, Carl and Wingren, Christer},
  issn         = {1876-4347},
  language     = {eng},
  number       = {5A},
  pages        = {503--513},
  publisher    = {Elsevier},
  series       = {New Biotechnology},
  title        = {Advancing the global proteome survey platform by using an oriented single chain antibody fragment immobilization approach.},
  url          = {http://dx.doi.org/10.1016/j.nbt.2015.12.001},
  volume       = {33},
  year         = {2016},
}