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.


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


  • Complement System Proteins