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NCL Method ITA-5.2

By Barry Neun, Edward Cedrone, Marina Dobrovolskaia

Analysis of Complement Activation by single-plex EIA or multiplex ELISA

Listed in Datasets | publication by group NCL Protocols

Version 2.0 - published on 09 Jul 2020 doi:10.17917/EQCT-3C51 - cite this

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Description

This document describes a protocol for quantitative determination of complement activation by an Enzyme Immunoassay (EIA).  The complement system represents an innate arm of immune defense and is named so because it “complements” the antibody-mediated immune response.  Three major pathways leading to complement activation have been described: they are the classical pathway, alternative pathway and lectin pathway (Figure 1).  The classical pathway is activated by immune (antigen-antibody) complexes.  Activation of the alternative pathway is antibody independent.  The lectin pathway is initiated by plasma protein mannose-binding lectin. 

The complement system is a group of ~30 protein that includes several components (C1 -C9), and Factors (B, D, H, I, and P).  Activation of any of the three pathways results in cleavage of the C3 component of the complement system (1, 2).

This protocol is intended for follow-up studies on samples which demonstrated a positive response in the qualitative assay (NCL method ITA 5.1). It can be performed as independent protocol when high throughput analysis is needed.

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