Antibodies reactive to cleaved sites in complement proteins enable highly specific measurement of soluble markers of complement activation. Issue 2 (August 2015)

• Record Type: Journal Article
• Authors: Blom, Anna M.; Österborg, Anders; Mollnes, Tom E.; Okroj, Marcin
• Is Part Of: Molecular immunology. Volume 66:Issue 2(2015:Aug.)
• Page Start: 164
• Page End: 170
• Publication Date: 2015-08
• Subjects: aa amino acid -- CAU complement activation unit -- FI complement factor I -- ICS#2 International Complement Standard #2 -- HRP horseradish peroxidase -- NHS normal human serum -- HI-NHS heat inactivated NHS -- OPD o-phenylenediamine dihydrochloride; Antibodies -- Complement system -- Leukemia -- Lymphoma -- C4d -- C5b; Immunochemistry -- Periodicals; Molecular biology -- Periodicals; Immunochemistry -- Periodicals; Allergy and Immunology -- Periodicals; Molecular Biology -- Periodicals; Immunochimie -- Périodiques; Biologie moléculaire -- Périodiques; Immunochemistry; Molecular biology; Periodicals; Electronic journals; 571.96
• DOI: 10.1016/j.molimm.2015.02.029
• Languages: English
• ISSNs: 0161-5890

Abstract

Highlights: We generated antibodies against 5 aa-long linear neoepitopes in C4d and C5b. The antibodies specifically recognize activated complement components. Measurement of C4d and TCC allows follow up of systemic complement activation. Abstract: An emerging number of diseases and therapeutic approaches with defined involvement of the complement system justify a need for specific markers reflecting activation of particular effector arms of the complement cascade. Measurement of such soluble markers in circulation is a challenge since the specificity of antibodies must be limited to activated complement fragments but not predominant and ubiquitous parental molecules. Existing assays for the measurement of soluble, activated complement proteins are based on the detection of conformational neoepitopes. We tested an alternative approach based on detection of short linear neoepitopes exposed at the cleavage sites after activation of the actual complement component. Obtained antibodies reactive to C4d and C5b fragments enabled us to set up highly specific sandwich ELISAs, which ensured trustful measurements without false positive readouts characteristic for some of the widely used assays.