Fundamental protein metrology to support the definition of measurands, analytical targets, and their associated measurement uncertainty

Proteins, which on a primary level consist of a sequence of amino acids, undergo complex folding mechanisms until they have reached their final structure. A protein’s function is highly correlated to its overall structure. However, during protein synthesis, modifications occur at various time points that can significantly alter the final structure of the protein. Therefore, a protein can be present in different versions called proteoforms. Protein quantitative measurements are essential for many medical tests, and therefore it is important to understand how the presence of proteoforms affects these measurements. This knowledge will further support the EU Global Health Strategy’s aim to improve the global health security across Europe. Specific metrology needs include investigating the influence of protein structure on measurements and developing accurate and SI-traceable higher order structure analysis protocols.

This project aims to develop an approach to identify and quantify the presence of proteoforms in heterogenous protein materials. Additionally, the project will publish protocols and guidelines for higher order protein structural analysis by cyclic ion mobility mass spectrometry (IM-MS), high-resolution cyclic IM-MS, and hydrogen deuterium exchange mass spectrometry. The project will also generate models to investigate the correlation between structure and function, as well as work on innovative in-vitro assays for proteins structure determination. These results will all be compiled in a metrology framework for protein characterisation methods, including the estimated measurement uncertainty values. These advancements will benefit the metrology community across Europe and support the United Nation’s sustainable development goal for good health and well-being.