Project Details
Projekt
Mass photometry platform
Subject Area
Basic Research in Biology and Medicine
Term
Funded in 2025
Project identifier
Deutsche Forschungsgemeinschaft (DFG) - Project number 559280663
The spectacular advances in understanding of biological macromolecules and their functional interactions are due to both technical developments in structural methods and novel approaches in sample preparation and characterization. Determining the dimensions of molecules based on their hydrodynamical radius with size exclusion chromatography and dynamic/static light scattering belongs to the fundamentals in biochemistry/structural biology. Major drawbacks of those methods are their limited ability to resolve the heterogeneity within the samples, e.g. discriminate and quantify monomeric and dimeric species due to the relatively small difference in the radius. Furthermore, the bulk methods are commonly linked with large sample consumption. Mass photometry is a recently developed method that uses light scattering at the single-molecule level to determine masses of particles present in solution. The contrast intensity from individual molecules present at low nanomolar concentration is recorded at the objective and converted then to the molecular weights. As multiple molecules are identified simultaneously, data acquisition typically takes one to several minutes, resulting in several hundreds to thousands of individual recognition events, which ensure sufficient statistics. Combination of the mass photometry with microfluidics belongs to the most recent advances, that allows performing the experiments at high, up to tens of micro-molar concentration of studied molecules. The sample is rapidly diluted when entering the measurement chamber followed by the immediate data acquisition, so transiently stable macromolecular complexes are resolved. The mass photometry is applicable to the broad range of macromolecules, such as proteins, nucleic acids, lipid-based nanoparticles etc., with the minimal mass of 30 kDa and up to 5 MDa, being the highly efficient cost- and time-wise method. Determination of the molecular weight and the homogeneity of sample is routinely required in biomolecular studies as a quality control step prior elaborate experiment, such as structural and biophysical analysis. Other applications of mass photometry include studying protein:protein interactions, where the assembled complex and its orphan components can be visualized in a straightforward way, or biochemical reactions, e.g. proteolysis, being a complementary tool for mass spectrometry-based proteomics. With a multiple of groups involved in biomolecular research in the University of Düsseldorf, a broad diversity of projects would greatly benefit from a microfluidics-based mass photometry platform.
DFG Programme
Major Research Instrumentation
Major Instrumentation
Mehrzweck-Massenphotometrie-Plattform
Instrumentation Group
5360 Meßgeräte für gestreutes und reflektiertes Licht, optische Oberflächen-Prüfgeräte
Applicant Institution
Heinrich-Heine-Universität Düsseldorf
