On-chip complement activation adds an extra dimension to antigen microarrays

Krisztián Papp, Zsuzsanna Szekeres, Nóra Terényi, Andrea Isaák, A. Erdei, J. Prechl

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Antibody profiling on antigen microarrays helps us in understanding the complexity of responses of the adaptive immune system. The technique, however, neglects another, evolutionarily more ancient apparatus, the complement system, which is capable of both recognizing and eliminating antigen and serves to provide innate defense for the organism while cooperating with antibodies on multiple levels. Complement components interact with both foreign substances and self molecules, including antibodies, and initiate a cascade of proteolytic cleavages that lead to the covalent attachment of complement components to molecules in nanometer proximity. By refining the conditions of antibody profiling on antigen arrays we made use of this molecular tagging to identify antigens that activate the complement system. Antigen arrays were incubated with serum under conditions that favor complement activation, and the deposited complement C3 fragments were detected by fluorescently labeled antibodies. We used genetically C3-deficient mice or inhibition of the complement cascade to prove that the technique requires complement activation for the binding of C3 to features of the array. We demonstrate that antigens on the array can initiate complement activation both by antibody-dependent or -independent ways. Using two-color detection, antibody and complement binding to the relevant spots was measured simultaneously. The effect of adjuvants on the quality of the immune response and binding of autoantibodies to DNA with concomitant complement activation in the serum of mice suffering from systemic autoimmune disease was readily measurable by this new method. We propose that measurement of complement deposition on antigen microarrays supplements information from antibody binding measurements and provides an extra, immune function-related fingerprint of the tested serum.

Original languageEnglish
Pages (from-to)133-140
Number of pages8
JournalMolecular and Cellular Proteomics
Volume6
Issue number1
DOIs
Publication statusPublished - Jan 2007

Fingerprint

Complement Activation
Microarrays
Chemical activation
Antigens
Antibodies
Serum
Complement C3
Molecules
Immune system
Dermatoglyphics
Autoantibodies
Autoimmune Diseases
Refining
Immune System
Color
DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

On-chip complement activation adds an extra dimension to antigen microarrays. / Papp, Krisztián; Szekeres, Zsuzsanna; Terényi, Nóra; Isaák, Andrea; Erdei, A.; Prechl, J.

In: Molecular and Cellular Proteomics, Vol. 6, No. 1, 01.2007, p. 133-140.

Research output: Contribution to journalArticle

Papp, Krisztián ; Szekeres, Zsuzsanna ; Terényi, Nóra ; Isaák, Andrea ; Erdei, A. ; Prechl, J. / On-chip complement activation adds an extra dimension to antigen microarrays. In: Molecular and Cellular Proteomics. 2007 ; Vol. 6, No. 1. pp. 133-140.
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