To design, construct, and run specific, sensitive, and robust assay is crucial to biomedical research. These assays play a crutial role in the pharmaceutical and biotechnology industries for the identification and characterization of potential new drugs.
Many factors influence the development of a biological test. Various bioanalytical assays are currently in use. Whatever may be the mechanism or method, the basic principles of assay development must be followed to get robust and reliable results to generate meaningful data.
Validation is establishing documented evidence that ensures that a specific assay will consistently produce a product or an outcome as per its predetermined specifications and quality attributes.
Parameters for Development and Validation of Robust assay:
When developing and validating a test for bioanalytical testing, the researcher should consider several parameters. These factors play a very important role in assay development. These parameters are:
- Nature of molecules to be measured
The researcher should be clear about the type and property of the molecules to be analyzed. All subsequent assay development activities will depend on the nature of the molecule under investigation.
- Source of the molecule
Information about the source of the molecule under analysis will determine the nature and quantity of the sample, its availability, concentration, and its stability. Hence, the source of the molecule has a significant influence on the assay development process.
Understanding the stability of the assay molecule is very important. Particles may be extremely stable in one matrix while unstable in another matrix.
- Number of samples
If only a small amount of samples is to be analyzed, a labor-intensive, multi-step manual assay format may prove to be suitable. But if the number of the run is more, it will be necessary to simplify and automate the assay process.
- Specificity, sensitivity, and range
The researcher needs to confirm that the assay will measure only the desired molecule and nothing else. The assay must be sufficiently sensitive so that the level and concentration of molecule falls well within the dynamic range of the assay
An assay procedure can produce results that are proportional to the level of the analyte in the sample.
- Reproducibility or Robustness
The assay must be robust and reproducible to give reliable and usable data. The article must be highly reproducible so that the degree of variation is negligible on every run. Robustness measures the capacity of an analytical procedure to remain unaffected by small but deliberate changes in method parameters and indicate its reliability. The evaluation of robustness is essential during the development phase and depends on the type of procedure under study.
If the process is susceptible to variations in analytical conditions, the analytical terms should be suitably under control, or a precautionary statement should be a part of the indications.
Assay development and validation of simple, sensitive, specific, and robust bioanalytical assays help biopharma companies and research laboratories in their new drug designing and development program and its subsequent use as therapeutics.