Heat Differentiated Complement Factor Profiling

J Proteomics. 2015 Aug 3;126:155-62. doi: 10.1016/j.jprot.2015.05.027. Epub 2015 Jun 3.

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.

Keywords: Complement factors; Dried blood spots; Immunodeficiency; Multiplex immunoassay; Neonatal screening; Serum.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Complement Activation*
  • Complement System Proteins / chemistry*
  • Complement System Proteins / deficiency
  • Complement System Proteins / metabolism
  • Female
  • Hot Temperature*
  • Humans
  • Male
  • Protein Stability

Substances

  • Complement System Proteins