What are the main optimization parameters for the Meso Scale Discovery assay?
Meso Scale Discovery is a platform to develop immunoassays that measure biomarkers in biological research and pre-clinical and clinical trials. MSD assay provides ultra-low analyte detection limits, uses a minute amount of sample, and is simple to perform.
Principle of MSD assay
It is a MultiArray technology that runs on the principle similar to ELISA. Specially designed microplates help create a novel assay for a single analyte, which can be multiplexed by using MSD MULTI-SPOT® plates. It increases the efficiency of the test.
MSD assays use electrochemiluminescent tags or labels in conjunction with antibodies to detect the light signal in the assay. These labels are known as SULFO-TAG and help in ultra-sensitive detection of the analyte.
MSD Assay optimization parameters
MSD assay Optimization involves individual testing of all the kit components and the complete kit’s measurement, considering it as a unit. The optimization parameters are the concentration of antibody, formulation of the buffer, formulation of the diluent, protocol optimization, preparation of sample and reagents, and scale-up of the manufacturing process.
Concentration of antibody
Antibody concentration is a critical parameter for any immunological assay. It is more so in the case of MSD as it measures the smallest amount of the analyte. The antigen-antibody complex formation depends upon the concentrations of both the reactants.
If the level of antibodies is not optimized, it may result in a situation called antibody access, which may hinder the formation of the antigen-antibody complex.
Formulation of the buffer
Buffer plays a vital role in an immunological assay. As antibodies are always protein in nature and antigen are mostly protein, a buffer helps resist the pH shift and thus protects the structure of these reactants.
Secondly, it helps in forming a suitable antigen-antibody complex, especially if the antigen is soluble. Hence, buffer formation must undergo proper optimization.
Formation of the diluent
Diluent, as the name indicates, has a function of diluting the concentration of reactants to bring it within the detection range of the assay. Optimization of the diluent is crucial for the sensitivity of the test.
The protocol of an assay indicates its entire process. Optimization of protocol affects the accuracy, efficacy, speed, and sensitivity of the test. The trial and error method or statistical methods can be of help in the optimization of these parameters.
Preparation of sample and reagents
Sample preparation is a very tricky but equally important process. The dissolution of the sample, sample dilution, concentration, and composition are all significant. Apart from the test sample, the preparation of other reagents for the assay is equally essential.
Optimization of reagent and sample preparation ensures the accuracy and sensitivity of the test.
Scale-up of the manufacturing process
It relates to the optimization of all the reagents and other components in a kit for the MSD assay. It also optimizes the quality of the reagents and other material in a kit.
Optimization of Meso Scale discovery assay concerning various parameters makes it more accurate and authentic. MSD assay makes diagnostics easy and fast.