Scaling Up Biopharmaceutical Characterization with the Orbitrap Ascend BioPharma Tribrid Mass Spectrometer

Biopharmaceutical pipelines are expanding rapidly, with increasingly diverse therapeutic modalities such as antibody-drug conjugates (ADCs), oligonucleotides, PROTACs, and RNA-based therapies. Each of these presents unique analytical challenges, from structural heterogeneity to low-abundance impurities, that require highly versatile and sensitive instrumentation.

Thermo Fisher Scientific: Scaling Up Biopharmaceutical Characterization with the Orbitrap Ascend BioPharma Tribrid Mass Spectrometer

The Thermo Scientific Orbitrap Ascend BioPharma Tribrid Mass Spectrometer was developed to meet these challenges head-on. Designed with a new dual ion routing multipole architecture, extended mass range detection, and integrated ease-of-use features, it enables comprehensive characterization of today’s most complex therapeutic molecules.

Scaling up insight for biopharma

The Orbitrap Ascend BioPharma MS introduces several key innovations that directly address bottlenecks in drug characterization:

• Native MS with extended mass range: Quadrupole isolation up to m/z 8,000 and Orbitrap detection up to m/z 16,000 allow detailed characterization of intact protein complexes and biotherapeutics in their native states.
• Proton Transfer Charge Reduction (PTCR): Simplifies highly heterogeneous spectra, enabling confident identification of proteoforms.
• Orthogonal fragmentation options (HCD, CID, ETD, EThcD, UVPD): Deliver near-complete sequence coverage and precise localization of post-translational modifications (PTMs) and conjugation sites.
• Ultra-high-field Orbitrap analyzer: Provides >480K resolution at FWHM and acquisition rates up to 45 Hz, supporting both depth and throughput.
• Auto-ready ion source: Enables automated, remote, and schedulable calibration, maintaining performance and consistency without interrupting experimental workflows.

Applications across the biopharma landscape

• Antibody-Drug Conjugates (ADCs)
  Middle-down analysis with PTCR and orthogonal fragmentation provides accurate drug-to-antibody ratio (DAR) determination and unambiguous localization of conjugation sites. Intact and fragmented analyses can be seamlessly combined in a single workflow.
• Host Cell Protein (HCP) monitoring
  The Orbitrap Ascend BioPharma MS detects trace HCPs at concentrations below 1 ppm at analytical flow rates, overcoming dynamic range challenges and supporting regulatory expectations for impurity profiling.
• PTM mapping
  Enhanced sensitivity enables the confident detection of low-abundance modifications (<1%), including challenging events such as deamidation, with 100% sequence coverage of both heavy and light antibody chains.
• Oligonucleotide and RNA therapeutics
  Low-q CID and UVPD fragmentation deliver nearly complete sequence coverage of intact oligonucleotides and sgRNAs, supporting the growing importance of gene and RNA-based therapies.
• Top-down proteomics
  The system’s versatility supports the direct characterization of intact proteoforms, providing clarity in structural biology and biopharmaceutical analysis.

Researcher perspective

According to Prof. Luca Fornelli, the University of Oklahoma:

“The Orbitrap Ascend BioPharma mass spectrometer combines two fundamental features that make it perfect for characterizing intact biopharmaceuticals. Multiple ion activation types are complemented by PTCR to simplify complex mass spectra and facilitate their interpretation. And the Native MS option increases the Orbitrap m/z range up to m/z 16,000. Together they enable researchers to perform sophisticated ion manipulations and analyze the most diverse molecules.”

Enabling confidence in therapeutic development

By scaling up analytical depth, throughput, and reproducibility, the Orbitrap Ascend BioPharma Tribrid Mass Spectrometer provides scientists with a powerful platform for characterizing complex therapeutics. Whether identifying trace-level impurities, localizing conjugation sites, or sequencing RNA molecules, Orbitrap Ascend MS delivers the clarity and confidence required to accelerate development timelines and meet regulatory expectations.

For Research Use Only. Not for use in diagnostic procedures.

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