Mass Photometry

Refeyn TwoMP mass photometer with MassFluidix® HC system

Room 6B (Sixth floor)
Contacts: Krzyszof Skowronek, Roman Szczepanowski
Collaboration/contracted research. Access to instrument  only by operator or authorized persons.

 

Refeyn TwoMP mass photometer

Mass photometry is a revolutionary new method of analyzing molecules. It enables the accurate mass measurements of single molecules in solution, in their native state and without the need for labels. For molecular mass measurements with unmatched sensitivity, speed and simplicity of use, a TwoMP mass photometer offers wide mass range and single-molecule resolution.

  • High-fidelity measurements of molecular mass

  • Minimal quantities of sample required

  • Intuitive acquisition and data analysis software 

  • Easy setup – a compact, benchtop instrument with minimum installation requirements

 

Protein-protein interactions

Antigen-antibody interactions are a prime example of molecular systems that can be studied using mass photometry, which can be applied to determine binding affinities for mono- and multivalent interactions.

Figure 1 illustrates how mass photometry can be
used to quantify molecular interactions. By measuring the mass of the antibody trastuzumab and its target antigen, Her2, individually and in mixtures, the interactions between individual antibody molecules and target antigens could be quantified.

DNA-protein interactions

Mass photometry can be used to characterise DNA-protein complexes, which are crucial in gene expression, replication and DNA repair.

In this example, mass photometry not only allows detection of DNA binding but also provides information on how the oligomeric state of the protein changes upon DNA binding (Figure 2).

Protein oligomerization

Many proteins adopt a particular oligomeric form under certain conditions. In Figure 1, mass photometry characterises four different proteins: protein A, beta-amylase, urease and thyroglobulin. Through this characterisation, these proteins showcase a range of behaviours – from purely monomeric (as in Protein A) to a dynamic equilibrium between multiple states.

Complex formation

Ribosomes are macromolecular assemblies of protein and RNA, and are central sites for protein synthesis. Bacterial ribosomes (in this example from E. coli) are >2 MDa in size. Magnesium ions play a subtle yet important role in the assembly of intact (70S) complexes. In the absence of magnesium, E. coli ribosomes rapidly disassemble into their 30S and 50S subunits. Mass photometry allows us to monitor these processes (Figure).

MassFluidix® HC system

Refeyn’s microfluidics system, MassFluidix HC, significantly expands the range of sample concentrations amenable to investigation by mass photometry, by raising the upper sample concentration limit from the nanomolar to the micromolar range. This enables applications such as the characterization of low-affinity interactions. 

MassFluidix HC is an add-on for Refeyn’s TwoMP mass photometer and includes a central unit with fluid controls and a rapid dilution chip.