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Development of Phage-Based Antibody Fragment Reagents for Affinity Enrichment of Bacterial Immunoglobulin G Binding Proteins.

Säll, Anna; Sjöholm, Kristoffer; Waldemarson, Sofia; Happonen, Lotta LU ; Karlsson, Christofer LU ; Persson, Helena and Malmström, Johan LU (2015) In Journal of Proteome Research 14(11). p.4704-4713
Abstract
Disease and death caused by bacterial infections are global health problems. Effective bacterial strategies are required to promote survival and proliferation within a human host, and it is important to explore how this adaption occurs. However, the detection and quantification of bacterial virulence factors in complex biological samples are technically demanding challenges. These can be addressed by combining targeted affinity enrichment of antibodies with the sensitivity of liquid chromatography-selected reaction monitoring mass spectrometry (LC-SRM MS). However, many virulence factors have evolved properties that make specific detection by conventional antibodies difficult. We here present an antibody format that is particularly well... (More)
Disease and death caused by bacterial infections are global health problems. Effective bacterial strategies are required to promote survival and proliferation within a human host, and it is important to explore how this adaption occurs. However, the detection and quantification of bacterial virulence factors in complex biological samples are technically demanding challenges. These can be addressed by combining targeted affinity enrichment of antibodies with the sensitivity of liquid chromatography-selected reaction monitoring mass spectrometry (LC-SRM MS). However, many virulence factors have evolved properties that make specific detection by conventional antibodies difficult. We here present an antibody format that is particularly well suited for detection and analysis of immunoglobulin G (IgG)-binding virulence factors. As proof of concept, we have generated single chain fragment variable (scFv) antibodies that specifically target the IgG-binding surface proteins M1 and H of Streptococcus pyogenes. The binding ability of the developed scFv is demonstrated against both recombinant soluble protein M1 and H as well as the intact surface proteins on a wild-type S. pyogenes strain. Additionally, the capacity of the developed scFv antibodies to enrich their target proteins from both simple and complex backgrounds, thereby allowing for detection and quantification with LC-SRM MS, was demonstrated. We have established a workflow that allows for affinity enrichment of bacterial virulence factors. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Proteome Research
volume
14
issue
11
pages
4704 - 4713
publisher
The American Chemical Society
external identifiers
  • pmid:26452057
  • wos:000364435100023
  • scopus:84946867028
ISSN
1535-3893
DOI
10.1021/acs.jproteome.5b00585
language
English
LU publication?
yes
id
c2eb92d9-4080-48b2-b4a5-945a7f6e33a9 (old id 8155585)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/26452057?dopt=Abstract
date added to LUP
2015-11-03 10:53:24
date last changed
2017-09-03 03:31:52
@article{c2eb92d9-4080-48b2-b4a5-945a7f6e33a9,
  abstract     = {Disease and death caused by bacterial infections are global health problems. Effective bacterial strategies are required to promote survival and proliferation within a human host, and it is important to explore how this adaption occurs. However, the detection and quantification of bacterial virulence factors in complex biological samples are technically demanding challenges. These can be addressed by combining targeted affinity enrichment of antibodies with the sensitivity of liquid chromatography-selected reaction monitoring mass spectrometry (LC-SRM MS). However, many virulence factors have evolved properties that make specific detection by conventional antibodies difficult. We here present an antibody format that is particularly well suited for detection and analysis of immunoglobulin G (IgG)-binding virulence factors. As proof of concept, we have generated single chain fragment variable (scFv) antibodies that specifically target the IgG-binding surface proteins M1 and H of Streptococcus pyogenes. The binding ability of the developed scFv is demonstrated against both recombinant soluble protein M1 and H as well as the intact surface proteins on a wild-type S. pyogenes strain. Additionally, the capacity of the developed scFv antibodies to enrich their target proteins from both simple and complex backgrounds, thereby allowing for detection and quantification with LC-SRM MS, was demonstrated. We have established a workflow that allows for affinity enrichment of bacterial virulence factors.},
  author       = {Säll, Anna and Sjöholm, Kristoffer and Waldemarson, Sofia and Happonen, Lotta and Karlsson, Christofer and Persson, Helena and Malmström, Johan},
  issn         = {1535-3893},
  language     = {eng},
  number       = {11},
  pages        = {4704--4713},
  publisher    = {The American Chemical Society},
  series       = {Journal of Proteome Research},
  title        = {Development of Phage-Based Antibody Fragment Reagents for Affinity Enrichment of Bacterial Immunoglobulin G Binding Proteins.},
  url          = {http://dx.doi.org/10.1021/acs.jproteome.5b00585},
  volume       = {14},
  year         = {2015},
}