Complete Overview of Assay Validation On Meso Scale Discovery (MSD) ELISA ECL Assay

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Meso Scale discovery (MSD) has a plethora of single and multiplex immunoassays for the detection and quantification of analyte in biological samples. With custom assay capabilities and over 400 assay kits, MSD has empowered researchers and scientists to accurately estimate phospholipids, cytokines, and other biomarkers in diverse biological matrices. meso scale discovery assays include single and multiplex options such as msdelisa, biomarker profiling assays, and a wide range of reagents for immunoassay development.

Unlike V-Plex MSD assays, which are validated MSD assays, MSD ELISA ECL assays should be validated before their use in the quantification of analytes. There are different aspects of assay validation that need to be addressed before running any immunoassay. Let us understand some of these crucial parameters that are necessary for the successful validation of an MSD ELISA ECL assay.

Overview of MSD ELISA ECL assay validation

Despite the popularity of multiplex immunoassays in basic and clinical research, few studies have specifically evaluated the pitfalls and challenges of commercially available kits across different clinical research settings. As mentioned earlier, other than V-Plex MSD assays, assays should be validated in their study settings.

MSD ECL platforms use a 96 well plate for analysis. These plates are similar to ELISA assays, but MSD plates are coated with electrodes to generate an electric impulse. Capture antibodies binding to the analyte of interest are placed at discrete spots in the wells. The detection antibodies comprise ECL labels that get activated through an electric charge. The activation only happens when the label is close to the electric charge. Hence almost all non-specific detection is eliminated.

The validation of an MSD ECL assay requires the testing of reproducibility, accuracy, and analyte recovery in test samples. To validate reproducibility, accuracy, and analyte recovery, it needs the demonstration of intra and inter-assay variability, and recovery and spike efficiency in the test assay matrix. Inter and intra assay variability are critical factors of a validated immunoassay. Intra-assay variability demonstrates the assay’s ability to repeat reliable results over different assays, whereas inter-assay variability is an assay’s ability to reproduce study results within the same assay.

Researchers add a known analyte in the test sample matrix and record its response by comparing it to a similar spike but in the standard diluent. Such spike and recovery data reveal whether the detection of an analyte is tampered with by the diluent used in preparing the biological sample matrix and standard curve. The generated data can then be compared to results obtained from traditional ELISA validation assays that are routinely used and previously validated. Such cross-validation ensures the reliability of the validated MSD assays. The use of validation methods encompassing reproducibility, accuracy, and analyte recovery is crucial before running any MSD ECL assay in study samples.

Final thoughts

Meso scale discovery electrochemiluminescence employs multi-array and electrochemiluminescence (ECL) detection capabilities for identifying biomarkers in both single and multiplex formats. ECL and multi-array technology facilitate speed and high density of research data through organization, miniaturization, and parallel processing of study samples. However, sponsors and research labs should invest in assay validation for ensuring the use of msd assays in routine research use.

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