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Onemp instrument

Manufactured by Refeyn
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The OneMP instrument is a compact and powerful mass photometer developed by Refeyn. It is designed to analyze the mass and size distribution of biological macromolecules, such as proteins, protein complexes, and nanoparticles, in solution. The OneMP instrument uses a unique optical detection method to provide accurate and reliable measurements without the need for labeling or additional sample preparation.

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Lab products found in correlation

Acquiremp, by Refeyn (6 mentions) Discovermp software, by Refeyn (4 mentions) 50 mm microscope coverslips, by Thermo Fisher Scientific (4 mentions) Gbl103250, by Merck Group (4 mentions) Nativemark unstained protein standard, by Thermo Fisher Scientific (4 mentions) Discovermp, by Refeyn (3 mentions) Lc0725, by Thermo Fisher Scientific (2 mentions) Acquiremp software, by Refeyn (2 mentions) Silicone gasket well sheet, by Grace Bio-Labs (2 mentions) Microscope coverslip, by Thermo Fisher Scientific (2 mentions) Superose 6 increase 10 300 gl, by Cytiva (1 mentions) Microscope coverslips, by Marienfeld (1 mentions) Thyroglobulin, by Merck Group (1 mentions) Gaskets, by Grace Bio-Labs (1 mentions) Beta amylase, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Protein standard, by Thermo Fisher Scientific (1 mentions) Twomp instrument, by Refeyn (1 mentions) Acquiremp v 2, by Refeyn (1 mentions)

Spelling variants of this product

OneMP (31 citations) OneMP mass photometer instrument (3 citations)

18 protocols using onemp instrument

1

Native Mass Spectrometry of Protein Complexes

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Samples were prepared using crosslinking at stoichiometric conditions, the reactions (±1.2 ml final volume after EDTA quenching) were concentrated to 500 μl and loaded on a pre-equilibrated Superose 6 10/300 increase GL (Cytiva) column in 20 mM HEPES–NaOH pH 7.5, 200 mM NaCl, 1 mM TCEP. Fractions were analyzed on SDS PAGE, the ones containing the complex of interest (Peak1 or Peak2) were pooled and concentrated to about 40 μl (Abs280 close to 0.5). The samples were diluted 10 to 20-fold in 20 mM HEPES–NaOH pH 7.5, 100 mM NaCl, 1 mM TCEP right before measuring on a Refeyn OneMP instrument (Refeyn Ltd). For each measurement, 13 μl of this buffer was first placed into the CultureWell gaskets wells (Grace Biolabs) placed into the Microscope coverslips (24 mm × 50 mm; Paul Marienfeld GmbH). After adjusting the focus, 2 μl of sample was mixed in. Movies were recorded for 60 seconds at 100 frames per second. A calibration measurement under the same conditions was performed roughly every 15 measurements using an in-house prepared protein standard mixture: IgG4Δhinge-L368A (73 kDa), IgG1-Campath (149 kDa), apoferritin (479 kDa), and GroEL (800 kDa). Data were processed using DiscoverMP (Refeyn Ltd) with bin width adjusted to 10, and each sample retrieved about 1500–3000 counts. Figures were prepared with the Refeyn instrument and edited in Illustrator.
Rouillon C., Eckhardt B.V., Kollenstart L., Gruss F., Verkennis A.E., Rondeel I., Krijger P.H., Ricci G., Biran A., van Laar T., Delvaux de Fenffe C.M., Luppens G., Albanese P., Sato K., Scheltema R.A., de Laat W., Knipscheer P., Dekker N.H., Groth A, & Mattiroli F. (2023). CAF-1 deposits newly synthesized histones during DNA replication using distinct mechanisms on the leading and lagging strands. Nucleic Acids Research, 51(8), 3770-3792.
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2

Mass Distribution Analysis of ASNase

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The mass distributions of ASNase samples were determined using a Refeyn OneMP instrument (Refeyn, Oxford, UK). Measurements followed the standard protocol [27 (link)]. Briefly, the sample chambers were assembled by placing silicon gasket wells on carefully selected and washed 24 × 50 mm coverslips as described in Wu and Piszczek [27 (link)]. Sample wells were partially filled with 10 μL of the standard buffer and filtered through 0.22 μm syringe filter. Enzyme solutions were cleared by centrifugation, concentrations were determined by the absorption at 280 nm and samples were diluted to concentrations of approximately 40 nM. Immediately before data acquisition 10 μL of the enzyme solutions were applied into the buffer-filled sample well and mixed by pipetting, resulting in 1 : 1 final dilution. A one-minute video was recorded using the AcquireMP software (Refeyn) and analysed using the DISCOVER MP software (Refeyn). In parallel to the measurements of enzymes, the mixture of known proteins (unstained protein ladder, LC0725, Thermo Fisher Scientific, Wattham, MA, USA) was measured under identical conditions to assure accurate calibration of molecular weights.
Strzelczyk P., Zhang D., Alexandratos J., Piszczek G., Wlodawer A, & Lubkowski J. (2022). The dimeric form of bacterial L-asparaginase YpAI is fully active. The FEBS journal, 290(3), 780-795.
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3

Nanodiscs for Protein Complex I

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The composition of the nanodisc-reconstituted protein preparation was assessed using mass photometry on a Refeyn OneMP instrument (Refeyn Ltd.), which was calibrated using an unstained native protein ladder (NativeMark Unstained Protein Standard A, Thermo Fisher Scientific Inc). Measurements were performed on the reconstituted complex I at a concentration of 0.015 mg ml−1 using AcquireMP 2.2.0 software and were analyzed using the DiscoverMP 2.2.0 package.
Kolata P, & Efremov R.G. (2021). Structure of Escherichia coli respiratory complex I reconstituted into lipid nanodiscs reveals an uncoupled conformation. eLife, 10, e68710.
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4

Nanobody Mass Profiling by MP

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Select nanobodies were binned using MP. Experiments were performed on a Refeyn OneMP instrument (Refeyn Ltd). The instrument was calibrated with a mix of BSA (Sigma-Aldrich), thyroglobulin (Sigma-Aldrich), and beta-amylase (Sigma-Aldrich). Coverslips (Thorlabs) and gaskets (Grace Bio-Labs) were prepared by washing with 100% IPA followed by ddH2O, repeated three times, followed by drying with HEPA filtered air. 12 μl of buffer was added to each well to focus the instrument after which 8 μl of protein solution was added and pipetted up and down to briefly mix after which movies/frame acquisition was promptly started. The final concentration in each experiment of recombinant spike S1 monomer (Sino Biological) and each nanobody was 30 nM and between 25 and 40 nM, respectively. Movies were acquired for 60 s (6000 frames) using AcquireMP (version 2.3.0; Refeyn Ltd) using standard settings. All movies were processed, analyzed, and masses estimated by fitting a Gaussian distribution to the data using DiscoverMP (version 2.3.0; Refeyn Ltd).
Mast F.D., Fridy P.C., Ketaren N.E., Wang J., Jacobs E.Y., Olivier J.P., Sanyal T., Molloy K.R., Schmidt F., Rutkowska M., Weisblum Y., Rich L.M., Vanderwall E.R., Dambrauskas N., Vigdorovich V., Keegan S., Jiler J.B., Stein M.E., Olinares P.D., Herlands L., Hatziioannou T., Sather D.N., Debley J.S., Fenyö D., Sali A., Bieniasz P.D., Aitchison J.D., Chait B.T, & Rout M.P. (2021). Highly synergistic combinations of nanobodies that target SARS-CoV-2 and are resistant to escape. eLife, 10, e73027.
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5

Oligomeric State Analysis of Proteins

Cited 1 time
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A Refeyn OneMP instrument (Refeyn Ltd) was used to measure the oligomeric states of the proteins in solution (45 (link)). Ten microliter of gel filtration buffer followed by 1 μl of the protein solution was applied to the drop to a final concentration of 100 nM and 6000 frames were recorded. The calibration curve was obtained by using three protein standards (66, 146, and 480 kDa) (Thermo Fisher Scientific).
Chua S.M., Wizrah M.S., Luo Z., Lim B.Y., Kappler U., Kobe B, & Fraser J.A. (2021). Structural features of Cryptococcus neoformans bifunctional GAR/AIR synthetase may present novel antifungal drug targets. The Journal of Biological Chemistry, 297(4), 101091.
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6

Purification and Characterization of Rel Proteins

Cited 3 times
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Detailed description of experimental procedures is provided in Supplementary methods. All bacterial strains and plasmids used in this study are listed in Supplementary Tables 1 and 2. All proteins were expressed, purified and characterized using SUMO-tagging strategy described earlier for E. coli RelA (33 (link)) and B. subtilis Rel (12 (link)) (Supplementary Figure S1A-G). The monomeric nature of 50 nM B. subtilis Rel and 100 nM E. coli RelA was confirmed by mass photometry (34 (link)) using Refeyn OneMP instrument (Refeyn Ltd.) (Supplementary Figure S1H and I).
Takada H., Roghanian M., Caballero-Montes J., Van Nerom K., Jimmy S., Kudrin P., Trebini F., Murayama R., Akanuma G., Garcia-Pino A, & Hauryliuk V. (2020). Ribosome association primes the stringent factor Rel for tRNA-dependent locking in the A-site and activation of (p)ppGpp synthesis. Nucleic Acids Research, 49(1), 444-457.
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7

Mass photometry analysis of C. crescentus DriD

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MP analysis was done using a Refeyn OneMP instrument (Oxford, UK). Contrast-to-mass calibration was achieved by measuring the contrast of proteins in the native marker protein standard mixture (NativeMark Unstained Protein Standard, Thermo Fisher) to generate a standard calibration curve. The experiments were performed using glass coverslips, which were thoroughly washed several times with Milli-Q water and isopropyl alcohol. Silicone gaskets were positioned on the glass surface for sample loading. FL C. crescentus DriD in 50 mM Tris pH 7.5, 300 mM NaCl, 5% glycerol was diluted in the buffer to final concentrations of 20 nM prior to sample analyses. 6000 frame movies were recorded using AcquireMP software and large field-of-view acquisition settings. The MP data was analyzed with DiscoverMP software to produce mass values.
Schumacher M.A., Cannistraci E., Salinas R., Lloyd D., Messner E, & Gozzi K. (2023). Structure of the WYL-domain containing transcription activator, DriD, in complex with ssDNA effector and DNA target site. Nucleic Acids Research, 52(3), 1435-1449.
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8

Mass Photometry Analysis of rAAV Samples

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The MP experiments were performed at room temperature using the OneMP instrument (Refeyn, Oxford, UK) following the standard protocol [35 (link)]. The 24×50 mm microscope coverslips (Fisher Scientific, Waltham, MA, USA) were prepared by cleaning with MilliQ water and isopropanol, and drying under a stream of clean nitrogen, as described previously [36 (link)]. A piece of clean, precut 2×2-well culturewell gasket (GBL103250, Sigma, MO, USA) was attached to the coverslip. The rAAV stocks’ concentrations were measured by the UV absorbance and diluted in PBS to a concentration of about 1011 virus particles per milliliter. This concentration was high enough to provide a high frequency of landing events without producing a large number of landing event overlaps that would degrade the data quality (Supplementary Table S1) [35 (link)]. All samples and stock solutions were carefully re-mixed just before use to assure sample homogeneity. Ten microliters of the filtered PBS buffer were loaded into a well of the culturewell gasket, and, after MP focusing, a 10 μl of rAAV sample was added into the same well. Immediately after the solution was mixed by pipetting, a 2 min video was recorded using the AcquireMP (Refeyn, Oxford, UK) software.
Wu D., Hwang P., Li T, & Piszczek G. (2022). Rapid characterization of adeno-associated virus (AAV) gene therapy vectors by mass photometry. Gene therapy, 29(12), 691-697.
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9

Mass Photometry Analysis of rAAV Samples

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The MP experiments were performed at room temperature using the OneMP instrument (Refeyn, Oxford, UK) following the standard protocol [35 (link)]. The 24×50 mm microscope coverslips (Fisher Scientific, Waltham, MA, USA) were prepared by cleaning with MilliQ water and isopropanol, and drying under a stream of clean nitrogen, as described previously [36 (link)]. A piece of clean, precut 2×2-well culturewell gasket (GBL103250, Sigma, MO, USA) was attached to the coverslip. The rAAV stocks’ concentrations were measured by the UV absorbance and diluted in PBS to a concentration of about 1011 virus particles per milliliter. This concentration was high enough to provide a high frequency of landing events without producing a large number of landing event overlaps that would degrade the data quality (Supplementary Table S1) [35 (link)]. All samples and stock solutions were carefully re-mixed just before use to assure sample homogeneity. Ten microliters of the filtered PBS buffer were loaded into a well of the culturewell gasket, and, after MP focusing, a 10 μl of rAAV sample was added into the same well. Immediately after the solution was mixed by pipetting, a 2 min video was recorded using the AcquireMP (Refeyn, Oxford, UK) software.
Wu D., Hwang P., Li T, & Piszczek G. (2022). Rapid characterization of adeno-associated virus (AAV) gene therapy vectors by mass photometry. Gene therapy, 29(12), 691-697.
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10

Biomass Quantification of Q-beta Conjugates

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The 24 × 50 mm microscope coverslips (Fisher Scientific, Waltham, MA) and precut 2 × 2 silicon gasket wells (GBL103250, Sigma, MO) were cleaned and assembled as described in the literature.46 (link) Measurements were performed on OneMP instrument (Refeyn, Oxford, UK) at room temperature. PBS (1×, pH 7.2) buffer was filtered through 0.22 μM filters before use. Ten microliters of the buffer were loaded in gasket well to focus the objective on the coverslip surface. Qβ conjugates stock were diluted 100 times in PBS (1×, pH 7.2) buffer and added to buffer in the well. The MP video was immediately recorded after the sample loading using the AcquireMP software (Refeyn, Oxford, UK). A 1 min video was recorded for each sample, and each sample was repeated twice. Data were processed using the DiscoverMP software (Refeyn, Oxford, UK) with the threshold filter values of 5. The mass distribution was plotted as histograms with bin width of 50 kDa and fit with Gaussian peaks to obtain the average mass of different species. The contrast-to-mass calibration was performed using an unstained protein ladder (LC0725, Thermo Fisher, Wattham, MA) and empty AAV5 sample (Virovek, Hayward, CA).
Rashidijahanabad Z., Kelly M., Kamruzzaman M., Qadri F., Bhuiyan T.R., McFall-Boegeman H., Wu D., Piszczek G., Xu P., Ryan E.T, & Huang X. (2022). Virus-like Particle Display of Vibrio cholerae O-Specific Polysaccharide as a Potential Vaccine against Cholera. ACS infectious diseases, 8(3), 574-583.
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