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Heat differentiated complement factor profiling.

Hamsten, Carl; Melander Skattum, Lillemor LU ; Truedsson, Lennart LU ; von Döbeln, Ulrika; Uhlén, Mathias; Schwenk, Jochen M; Hammarström, Lennart; Nilsson, Peter and Neiman, Maja (2015) In Journal of Proteomics 126. p.155-162
Abstract
Complement components and their cascade of reactions are important defense mechanisms within both innate and adaptive immunity. Many complement deficient patients still remain undiagnosed because of a lack of high throughput screening tools. Aiming towards neonatal proteome screening for immunodeficiencies, we used a multiplex profiling approach with antibody bead arrays to measure 9 complement proteins in serum and dried blood spots. Several complement components have been described as heat sensitive, thus their heat-dependent detectability was investigated. Using sera from 16 patients with complement deficiencies and 23 controls, we confirmed that the proteins C1q, C2, C3, C6, C9 and factor H were positively affected by heating, thus the... (More)
Complement components and their cascade of reactions are important defense mechanisms within both innate and adaptive immunity. Many complement deficient patients still remain undiagnosed because of a lack of high throughput screening tools. Aiming towards neonatal proteome screening for immunodeficiencies, we used a multiplex profiling approach with antibody bead arrays to measure 9 complement proteins in serum and dried blood spots. Several complement components have been described as heat sensitive, thus their heat-dependent detectability was investigated. Using sera from 16 patients with complement deficiencies and 23 controls, we confirmed that the proteins C1q, C2, C3, C6, C9 and factor H were positively affected by heating, thus the identification of deficient patients was improved when preheating samples. Measurements of C7, C8 and factor I were negatively affected by heating and non-heated samples should be used in analysis of these components. In addition, a proof of concept study demonstrated the feasibility of labeling eluates from dried blood spots to perform a subsequent correct classification of C2-deficiencies. Our study demonstrates the potential of using multiplexed single binder assays for screening of complement components that open possibilities to expand such analysis to other forms of deficiencies. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Proteomics
volume
126
pages
155 - 162
publisher
Elsevier
external identifiers
  • pmid:26047714
  • wos:000359884600014
  • scopus:84930942972
ISSN
1874-3919
DOI
10.1016/j.jprot.2015.05.027
language
English
LU publication?
yes
id
eff8af78-0962-4f22-80c9-d64abaf846ae (old id 7488043)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/26047714?dopt=Abstract
date added to LUP
2015-07-07 11:28:27
date last changed
2017-01-22 03:18:47
@article{eff8af78-0962-4f22-80c9-d64abaf846ae,
  abstract     = {Complement components and their cascade of reactions are important defense mechanisms within both innate and adaptive immunity. Many complement deficient patients still remain undiagnosed because of a lack of high throughput screening tools. Aiming towards neonatal proteome screening for immunodeficiencies, we used a multiplex profiling approach with antibody bead arrays to measure 9 complement proteins in serum and dried blood spots. Several complement components have been described as heat sensitive, thus their heat-dependent detectability was investigated. Using sera from 16 patients with complement deficiencies and 23 controls, we confirmed that the proteins C1q, C2, C3, C6, C9 and factor H were positively affected by heating, thus the identification of deficient patients was improved when preheating samples. Measurements of C7, C8 and factor I were negatively affected by heating and non-heated samples should be used in analysis of these components. In addition, a proof of concept study demonstrated the feasibility of labeling eluates from dried blood spots to perform a subsequent correct classification of C2-deficiencies. Our study demonstrates the potential of using multiplexed single binder assays for screening of complement components that open possibilities to expand such analysis to other forms of deficiencies.},
  author       = {Hamsten, Carl and Melander Skattum, Lillemor and Truedsson, Lennart and von Döbeln, Ulrika and Uhlén, Mathias and Schwenk, Jochen M and Hammarström, Lennart and Nilsson, Peter and Neiman, Maja},
  issn         = {1874-3919},
  language     = {eng},
  pages        = {155--162},
  publisher    = {Elsevier},
  series       = {Journal of Proteomics},
  title        = {Heat differentiated complement factor profiling.},
  url          = {http://dx.doi.org/10.1016/j.jprot.2015.05.027},
  volume       = {126},
  year         = {2015},
}