Paper detail

Importance of method validation: Implications of non-correlated observables in sweet taste receptor studies

Validation of research methodology is critical in research design. Correlation between experimental observables must be established before undertaking extensive experiments or propose mechanisms. This article shows that, observables in the popular calcium flux strength assay used in the characterization of sweetener-sweet taste receptor (STR) interaction are uncorrelated. In pursuit to find potential sweeteners and enhancers, calcium flux generated via G-protein coupling for wildtype and mutant receptors expressed on cell surface is measured to identify and localize sweetener binding sites. Results are channeled for sweetener development with direct impact on public health. We show that flux strength is independent of EC50 and sweet potency. Sweet potency-EC50 relation is non-linear and anti-correlated. Single point mutants affecting receptor efficiency, without significant shift in EC50 have been published, indicating flux strength is independent of ligand binding. G-protein coupling step is likely observed in the assay. Thus, years have been spent generating uncorrelated data. Data from uncorrelated observables does not give meaningful results. Still, majority of research in the field, uses change in calcium flux strength to study the receptor. Methodology away from flux strength monitor is required for sweetener development, reestablish binding localization of sweeteners established by flux strength method. This article serves to remind researchers to validate methodology before plunging into long term projects. Ignoring validation test on methodology, have been a costly mistake in the field. Concepts discussed here is applicable, whenever observable in biological systems are many steps moved from the event of interest.