Get deep structural insight to complex proteins and protein interactions
The complexity of modern therapeutic proteins, such as antibody-drug conjugates (ADCs), presents a significant analytical challenge requiring high resolution chromatography combined with high resolution mass spectrometry. ADCs involving cysteine conjugation present a unique challenge for characterization as proper intact analysis requires native mass spectrometry conditions to preserve structurally-critical non-covalent binding between antibody chains. Intact protein analysis under native mass spectrometric conditions can now be performed routinely using mass spec compatible buffers together with size exclusion (SEC) or ion exchange chromatography (IEC) coupled to Q Exactive BioPharma series mass spectrometers.
Native intact mass analysis workflow
Featured chromatography solutions for native intact mass
Use Thermo Scientific MAbPac SEC-1 Size Exclusion Columns for high-resolution separation of monoclonal antibody (mAb) analysis, including monomers and aggregates. These size-exclusion chromatography (SEC) columns provide superior performance, under non-denaturing conditions, and in high- and low-salt mobile phases and volatile eluents.
Use the innovative architecture of a UHPLC system that has been tailored to the specific and demanding analytical requirements that these essential drugs impose. The Thermo Scientific Vanquish Flex Binary and Quaternary systems have been designed and manufactured to exacting engineering standards to be truly built for biopharma.
Orbitrap mass spectrometers for native intact mass analysis
| Orbitrap Exploris 480 Mass Spectrometer | Orbitrap Eclipse Tribrid Mass Spectrometer | Q Exactive UHMR | Q Exactive BioPharma Platform | |
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| Advantages | Extraordinary simplified within a compact footprint where next level exploration delivers richer insights gained with intact protein analysis under native mass spectrometric conditions, empowering all by bringing performance and intelligence together. | 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 with intact protein analysis under native mass spectrometric conditions. | Go beyond what you thought possible with an outstanding tool for investigating the higher order structure, native-like tertiary and quaternary protein structures with a high resolution accurate mass and an ability to analyze ultrahigh mass range. | Realize complete BioPharma characterization with distinct operational modes optimized for the top three protein characterization workflows and a high mass range (HMR) mode for native MS analysis ideal for antibody drug conjugates and other heterogeneous antibody samples. |
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| Resolving power | 480,000 at m/z 195 | 7,500-500,000 FWHM at m/z 200 (1M option) | 200,000 FWHM at m/z 400 (1M option) | Up to 240,000 at m/z 200 |
| Scan speed | Up to 40Hz | Up to 40Hz | 12Hz at resolution setting of 12,500 at m/z 400 | 18Hz |
| 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) | 350 to 80,000 | 50 to 8,000 m/z |
| Dynamic range | >5000:1 | >5000 with a single OTMS spectrum | >5,000 with a single OTMS 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 successfully prevents interferences from co-eluting compounds and increases signal to noise ratios by 100-fold or higher.
Featured software for native intact mass analysis
BioPharma Finder integrated software provides confident protein deconvolution to determine the molecular weight of proteins in both denaturing and native conditions. Protein sequence matching identifies N-linked glycosylation and other common post-translational modifications (PTMs) via intact mass analysis, helping you to fully understand the heterogeneity of your biologic drug.
Three ways to spend less and get more
Upgrade your ability to achieve new scientific discoveries faster with Thermo Scientific Orbitrap mass analyzers that set new standards in sensitivity, performance and productivity. Or take advantage of upgrades to extend your current Thermo Scientific LC-MS system capabilities.
Watch videos on native intact mass analysis
Join experts from Genentech and Thermo Fisher to get great structural insight into your drug product using the Q Exactive BioPharma platform of mass spectrometers.
