Intact non-reduced antibody characterization

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 Thermo Scientific Orbitrap Exploris BioPharma series mass spectrometers.

Contact us  Join our community

On this page

Native intact mass analysis workflow

Featured chromatography solutions for native intact mass

Superior separations under non-denaturing conditions

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.

Chromatography built for biopharma

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 SpectrometerOrbitrap Eclipse Tribrid Mass SpectrometerQ Exactive UHMROrbitrap
Exploris 240 MS
 Orbitrap Exploris 480 Mass SpectrometerOrbitrap Eclipse Tribrid Mass SpectrometerOrbitrap Fusion Lumos Tribrid Mass SpectrometerOrbitrap Exploris 240
AdvantagesExtraordinary 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 protein analysis, and take advantage of the extended mass range up to m/z 8000 available with the BioPharma Option for native MS analysis for antibody drug conjugates and other heterogeneous antibody samples.
Resolving power480,000 at m/z 1957,500-500,000 FWHM at m/z 200 (1M option)200,000 FWHM at m/z 400 (1M option)Up to 240,000 FWHM at m/z 200
Scan speedUp to 40HzUp to 40Hz12Hz at resolution setting of 12,500 at m/z 400Up to 22Hz at resolution setting 15,000 at m/z 200
Mass range40 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 80,00040 to 8,000 m/z
Dynamic range>5000:1>5000 with a single OTMS spectrum>5,000 with a single OTMS spectrum>5000 within a single Orbitrap mass analyzer spectrum
Mass accuracyInternal <1 ppm RMS; External: <3 ppm RMSInternal <1 ppm RMS; External: <3 ppm RMSInternal <1 ppm RMS; External: <3 ppm RMSInternal <1 ppm RMS; External: <3 ppm RMS

Boost selectivity and enhance your proteomics workflow with FAIMS Pro Interface

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.

Learn more  Watch free webinar

Featured software for native intact mass analysis

Powerful protein deconvolution algorithms

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.

Watch videos on native intact mass analysis



Case studies

Application notes & posters