Antibodies reactive to cleaved sites in complement proteins enable highly specific measurement of soluble markers of complement activation.

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Antibodies reactive to cleaved sites in complement proteins enable highly specific measurement of soluble markers of complement activation. / Blom, Anna; Österborg, Anders; Mollnes, Tom E; Okroj, Marcin.

In: Molecular Immunology, Vol. 66, No. 2, 2015, p. 164-170.

Research output: Contribution to journalArticle

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TY - JOUR

T1 - Antibodies reactive to cleaved sites in complement proteins enable highly specific measurement of soluble markers of complement activation.

AU - Blom, Anna

AU - Österborg, Anders

AU - Mollnes, Tom E

AU - Okroj, Marcin

PY - 2015

Y1 - 2015

N2 - 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.

AB - 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.

U2 - 10.1016/j.molimm.2015.02.029

DO - 10.1016/j.molimm.2015.02.029

M3 - Article

VL - 66

SP - 164

EP - 170

JO - Immunochemistry

T2 - Immunochemistry

JF - Immunochemistry

SN - 1872-9142

IS - 2

ER -