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How we measure and analyze complement activation

We recommend that a uniform protocol is used for:
> Standardized sample collection
> Shipment
> Pre-analytical handling

> Storage methods for both the assessment of complement function as well as the measurements of individual complement components.

Study design

A more detailed description of the study design is needed regarding used methods and materials (e.g., type of collection tubes). For functional testing, complement-preserved serum (prepared as indicated in A) should be used. To assess individual complement components, EDTA-plasma must be used to minimize artificial complement activation. A first set-up for this uniform protocol is outlined in A

Source; Pubmed.ncbi

Standardization of complement analysis

The assessment itself should be performed using standardized assays to produce results that are robust and comparable between diagnostic laboratories. Regarding the assays, standardized calibrators and reagents must be used. Antibody-based assays should preferably use antibodies recognizing the same epitope to minimize the variation in kinetics of the antibodies for the antigen.

A flowchart for the selection of the appropriate technique for the assessment of complement is shown in B.

Standardization of complement analysis is organized and coordinated by the Sub-Committee for the Standardization and Quality Assessment of Complement Measurements (the ‘Complement EQA Group’). We believe that this group should be strongly embedded within and supported by the complement society so that they can have a leading role in providing clear directives for complement measurements.

Source; Pubmed.ncbi

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