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.

Rapid determination of stability by monitoring aggregation and degradation in a single measurement

High-fidelity measurements of molecular mass

Intuitive acquisition and data analysis software

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

With its high sensitivity, the TwoMP is ideally suited for measurements at physiological (i.e. low) concentrations. The high dynamic range intrinsic to single molecule counting techniques ensures low-abundance species are still captured accurately.

Refeyn’s software automatically controls the acquisition process and performs the mass analysis in minutes. You can intuitively interpret the mass distribution results without the need for any prior knowledge.

A compact, bench-top instrument with minimal installation requirements. Measurements only require a few microliters of sample and clean sample-carrier slides.

TwoMP Auto | Reliable, accurate walkaway solution

Automated mass photometry combines the ease and efficiency of automation with the unmatched sensitivity and simplicity of Refeyn’s molecular mass measurement technology. The automated mass photometer frees up operator time and offers enhanced precision, enabling rapid measurement of multiple samples with low sample consumption. Automation is compatible with the TwoMP.

Rapid, automated measurement of multiple samples

Ideal for screening and titration assays

Highly reproducible data with a user-friendly software

The TwoMP Auto streamlines your workflow, with highly reproducible data & a user-friendly software

'With the TwoMP Auto, you can run 24 measurements in as little as 90 minutes (or up to 180 minutes, depending on the protocol).

The TwoMP Auto delivers automated mass photometry

Start where you are today and scale as you grow – both configurations deliver the same trusted mass photometry results.

• Work with antibodies or other proteins, or nucleic acids • Require the highest mass resolution available in mass photometry • Want to deeply characterize sample purity and homogeneity, aggregation, binding, molecular integrity, stoichiometry and complex formation • Require full access to raw data for custom analysis • Need publication-ready figures that can be fully customized • Want to use application-specific software packages from Refeyn

• Need all the capabilities of TwoMP with automated sample handling for higher throughput • Need to screen multiple samples or conditions in a single, unattended session • Want significant walkaway time and consistent, reproducible results across samples and operators

• Have a TwoMP mass photometer • Work with proteins • Need to measure samples at higher concentrations than standard TwoMP allows (up to tens of micromolar) • Want to detect and characterize weak or transient interactions • Require rapid, minimal-sample analysis with quick experimental setup

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.

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.

The liquid-handling robot autonomously runs 24 measurements in *as little as 90 minutes (up to 180 minutes, depending on the protocol).

A straightforward, repeated measurement of a sample containing the antibody IgG illustrates the precision of the automated mass photometer, which is reflected in the high degree of reproducibility of the resulting data (Fig. 1).

Figure 1A shows histograms of 12 repeated measurements of an IgG sample made using automated mass photometry. Across the wells that contain the sample, the measured mass (Fig. 1B) and total counts (Fig. 1C) show low variability.

Figure 1 Automated mass photometry measurements are highly reproducible. Autonomously run mass photometry measurements show (A) the mass distributions of IgG forming monomers and dimers (inset: mass histograms for each well). The measured mass (B) and total counts (C) across the wells show low variability.

Q: What type of biomolecules can I measure with the TwoMP? A: The TwoMP has a wide mass range, allowing the study of different biomolecules, including proteins and protein complexes, nucleic acids, and large macromolecular complexes.

Q: What are the optimization steps necessary for a mass photometry measurement? A: The only optimization step necessary prior to a mass photometry measurement is to adjust the final concentration of the sample; the sample concentration on the instrument should be between 100 pM and 100 nM. The optimal concentration range for a mass photometry measurement is 5-20 nM.

Q: Does the shape/conformation of my protein affect the mass measured? A: For biomolecules within the mass detection range of the TwoMP (30 kDa to 5 MDa), the shape/conformation doesn’t influence the interferometric contrast and thus mass measured by the mass photometer.

Q: If my protein of interest is fluorescent under the laser light, will it affect the mass measured? A: In mass photometry, the mass measurement depends on the interference between the light scattered by the molecule and reflected light from the glass surface. This interference signal will be orders of magnitude greater than the fluorescence signal, so the mass measurement will not be affected.

Q: Which mass photometers are compatible with the ‘Auto’? A: If you already have a TwoMP, and SamuxMP, these mass photometers can be upgraded to automated systems. New automated mass photometers are available for purchase as TwoMP Auto and SamuxMP Auto.

Q: What consumables are available for the TwoMP Auto? A: We recommend using the Refeyn mass photometry consumables for Automated Systems. If you have a new TwoMP Auto instrument, you can order a Starter Kit, which includes everything you need to prepare samples and run a measurement (sample carrier slides and sample well cassettes to support 120 sample measurements, and the Alignment Assist Package to affix the sample well cassette). You can top up your supply of consumables with the Sample Preparation Package, which includes ready-to-use sample carrier slides and sample well cassettes to support 300 sample measurements or the Sample Carrier Slide-Only Package for Automated Systems (based on your needs).

Q: Is the TwoMP Auto able to dilute samples? A: Yes, there is an ‘in-plate’ dilution capability, and the user can select the following options in the software (version R1 2023): 1/2, 1/4, 1/8, 1/10, 1/25, 1/50, 1/100, 1/250, 1/500, 1/1000.

Q: What are the specs for TwoMP Auto? A: In terms of mass photometry, these are the same as the mass photometer inside the ‘Auto’. For ‘Auto’ specific specs, please refer to the data sheet.

Key Features

• 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
• High-fidelity measurements: With its high sensitivity, the TwoMP is ideally suited for measurements at physiological (i.e. low) concentrations. The high dynamic range intrinsic to single molecule counting techniques ensures low-abundance species are still captured accurately.
• Intuitive & efficient: Refeyn’s software automatically controls the acquisition process and performs the mass analysis in minutes. You can intuitively interpret the mass distribution results without the need for any prior knowledge.
• Minimal setup required: A compact, bench-top instrument with minimal installation requirements. Measurements only require a few microliters of sample and clean sample-carrier slides.
• Rapid, automated measurement of multiple samples
• Highly reproducible data with a user-friendly software
• Compatible with TwoMP unit
• The TwoMP Auto streamlines your workflow, with highly reproducible data & a user-friendly software
• The TwoMP Auto delivers automated mass photometry
• Automated mass characterization: A user-friendly software interface; Automation unit compatible with TwoMP; Ideal for screening and titration assays
• Highly reproducible data: Precision pipetting; Consistent measurements
• Intuitive operation: A user-friendly software interface; Automation unit compatible with TwoMP
• Highly reproducible results: The liquid-handling robot autonomously runs 24 measurements in *as little as 90 minutes (up to 180 minutes, depending on the protocol). A straightforward, repeated measurement of a sample containing the antibody IgG illustrates the precision of the automated mass photometer, which is reflected in the high degree of reproducibility of the resulting data (Fig. 1). Figure 1A shows histograms of 12 repeated measurements of an IgG sample made using automated mass photometry. Across the wells that contain the sample, the measured mass (Fig. 1B) and total counts (Fig. 1C) show low variability.

Applications

• 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.
• 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.

Specifications

TwoMP
Mass detection range	30 kDa to 5 MDa
Sample concentration on instrument	100 pM to 100 nM
Sample volume	a few microliters of sample
TwoMP Auto
Throughput	24 measurements in as little as 90 minutes (up to 180 minutes, depending on the protocol)
In-plate dilution options (software R1 2023)	1/2, 1/4, 1/8, 1/10, 1/25, 1/50, 1/100, 1/250, 1/500, 1/1000
TwoMP MassFluidix HC add-on
Sample concentration	up to tens of micromolar
TwoMP Auto
Compatible instrument	TwoMP (existing TwoMP and SamuxMP units can be upgraded)

Documents

• TwoMP Auto product data sheet
• MassFerence P1 product data sheet
• Refeyn Bioanalytics With Mass Photometry (brochure)

Related Products

• MassFluidix™ HC – Sample Preparation Plate
• MassGlass™ UC – Sample Prep Kit
• MP – Alignment Kit
• MassFerence™ P1 Calibrant
• MassFluidix™ HC – Sample Prep Kit