Deliver unequivocal intact mass assignments
Among the analytical tools used for the characterization of proteins, high-resolution accurate-mass (HRAM) mass spectrometry has become increasingly important for quantification of the various glycoforms and protein variant analysis. New methods coupling UHPLC with Orbitrap-based mass spectrometers give rapid insight into the proteoform profile for direct comparison of biotherapeutic products during discovery, development, and QC.
Intact protein mass spectrometry workflow
Featured chromatography solutions for intact mass analysis
Thermo Scientific Vanquish Flex UHPLC delivers confidence, with a fully bio-compatible flow path and proven compliance. Achieve unprecedented performance in retention time stability, sensitivity and separation efficiency with the widest range of column chemistries for biotherapeutic protein characterization. Demand versatility through seamless integration with market-leading mass spectrometry, fluorescence and charged aerosol detection. Gain operational simplicity with easy, freely available, one-click workflows via AppsLab and tool-free Viper fittings.
Perform high-performance, reversed-phase (RP) LC and LC-MS characterization of monoclonal antibodies (mAbs), fragments, variants, antibody drug conjugates (ADCs), PEGylated proteins, and bispecific proteins using the new MAbPac RP columns. These columns have excellent resolution, long column lifetime, and low carry over. The surface chemistry, particle size, and pore size, have been optimized to for high-resolution separations.
Reduce background contamination and noise seen in LC-MS analysis of samples containing non-volatile salts with a MSPac DS-10 desalting cartridge. Removal of these salts prior to analysis enhances analyte response and improves sensitivity aiding a more reliable identification. Use alone or in combination with an analytical column for high resolution separations.
Orbitrap mass spectrometers for intact mass analysis
| Orbitrap Exploris 480 Mass Spectrometer | Orbitrap Eclipse Tribrid Mass Spectrometer | Orbitrap Exploris 240 Mass Spectrometer | Q Exactive UHMR Mass Spectrometer | |
|---|---|---|---|---|
 |  |  |  | |
| Advantages | Extraordinary simplified within a compact footprint where next level exploration delivers quantifiable accuracy and the selectivity required to obtain richer insights, empowering all by bringing performance and intelligence together to provide rapid confirmation of intact protein molecular weight and determine glycoform heterogeneity of biopharmaceuticals. | Go beyond today's discovery with new innovations that deliver the ultimate flexibility to expand experimental scope, with built-in intelligence to ensure the highest data quality and confidence when confirming intact protein molecular weight and determining glycoform heterogeneity of biopharmaceuticals. | Superb spectral clarity delivers high-confidence biotherapeutic characterization to reliably quantitate and identify biotherapeutics in both research and routine laboratory environments. From intact mass analysis under denaturing and native conditions, to subunit analysis, middle- and top-down analysis, and peptide or disulfide bond mapping. | With orders of magnitude enhanced sensitivity at high m/z and its superior ability to perform native intact mass and top-down analyses leveraging ultra-high mass quadrupole selection up to m/z 25k, it provides unprecedented structural detail you can't see with other methods, enabling a deeper understanding of complex biological processes. |
| Resources | ||||
| Resolving power | 480,000 at m/z 195 | 7,500-500,000 FWHM at m/z 200 (1M option) | Up to 240,000 FWHM at m/z 200 | Resolution up to 200,000 FWHM at m/z 400 |
| Scan speed | Up to 40Hz | Up to 40Hz | 22Hz | 12 Hz at resolution setting of 12,500 at m/z 400 |
| Mass range | 40 to 6,000 m/z (up to 8,000 m/z with the BioPharma option) | 50 to 6,000 (up to 8,000 with HMRn) | 40 to 8,000 m/z | Ultra-high mass range up to m/z 80,000 |
| Dynamic range | >5000:1 | >5000 with a single OTMS spectrum | >5000 within a single Orbitrap mass analyzer spectrum | >5000:1 |
| Mass accuracy | Internal <1 ppm RMS; External: <3 ppm RMS | Internal <1 ppm RMS; External: <3 ppm RMS | Internal <1 ppm RMS; External: <3 ppm RMS | Internal: <1 ppm RMS External: <3 ppm RMS |
High-Field Asymmetric waveform Ion Mobility Spectrometry (FAIMS) can be used to optimize ion selectivity and reduce chemical backgrounds during mass spectrometry assays. FAIMS selectivity successfully prevents interferences from co-eluting compounds and increases signal to noise ratios by 100-fold or higher.
Featured software for intact mass analysis
Screen, identify, and characterize intact proteins with higher productivity and confidence using the Intact Protein workflow in BioPharma Finder software. Two deconvolution algorithms take full advantage of the high-quality HRAM data produced by Orbitrap mass spectrometers. In addition to intact characterization, you can confirm amino acid sequence, identify site and type of known/unknown post translational modifications, while providing relative amounts using Peptide Mapping workflows. Additional features include disulfide linkage, sequence variant analysis, and sequence alterations.
Featured videos
Join experts from Genentech and Thermo Fisher to get great structural insight into your drug product using Orbitrap high resolution accurate mass spectrometers.