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Workpackage 3: Assay development: transcriptome

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Objectives
  • To develop colorimetric immunoassays (ELISAs) for a panel of biomarkers defined in Work Package 1, which are believed to have the potential of predictive value with respect to cardiovascular risk.
  • To prepare reagents to allow the resulting ELISAs to be evaluated on a case cohort panel of ~11K samples
  • To transfer the ELISA methodology into the laboratory carrying out the testing (Universitätsklinikum Hamburg-Eppendorf, UKE)
  • Biomarker panel: Serpin F2, ABCA-1, ABCG-1, GPCR-15

Description of Work

For each of the selected biomarkers, it is required to identify suitable immunoreactive reagents (antibodies, antigens) and to use these to establish enzyme-based immunoassays which allow sensitive detection of the biomarker in human samples. This may involve screening a range of antibodies and antigens from commercial sources, or the generation of novel antibodies or antigens.
The effectiveness of each ELISA for its purpose can be validated by using it to detect the biomarker in samples provided by other consortium members, where there is known or expected concentration of the biomarker present.

Once ELISA designs have been established and their effectiveness verified, reagents will be prepared (~300 x 96-well microtitre plates plus associated liquid reagents, etc.) to allow UKE to evaluate ~11K case cohort samples, and a transfer process will be executed to ensure that UKE can use the ELISA effectively.

In the event that the biomarker shows promise with the case cohort samples, it may be decided to test a wider range of samples, in which case reagents will be prepared to allow this exercise too.


Partners involved

Fleet Bioprocessing (WP coordinator), Cavadis Utrecht, UMC Utrecht, UKE Hamburg