Receptor-binding hydrogen–deuterium exchange mass spectrometry as an additional measurement of biosimilarity

Treuheit, Nicholas A.; Crawford, Nicholas F.; Maki, Steven L.; Payne, Jason W.; Allen, Jeff

doi:10.1007/s40005-019-00465-9

Cited by 1 publication

(2 citation statements)

References 31 publications

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“…Bottom-up protein studies based on peptide mass mapping and peptide sequencing of filgrastim reported an a.a. sequence coverage up to 98.9% . It should be noted that sequence coverage is calculated differently in top-down and bottom-up strategies.…”

Section: Resultsmentioning

confidence: 99%

“…Bottom-up protein studies based on peptide mass mapping and peptide sequencing of filgrastim reported an a.a. sequence coverage up to 98.9%. 25 It should be noted that sequence coverage is calculated differently in top-down and bottom-up strategies. In bottom-up methods, all residues included in identified peptides are counted, regardless of the completeness of the fragment coverage within the identified peptide.…”

Section: Contribution Of Individual Ms/ms Fragmentation Modes To the ...mentioning

confidence: 92%

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Effective Amino Acid Sequencing of Intact Filgrastim by Multimodal Mass Spectrometry with Topdownr

Babović

Shliaha

Gibb

et al. 2022

J. Am. Soc. Mass Spectrom.

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Therapeutic proteins, known as biologicals, are an important and growing class of drugs for treatment of a series of human ailments. Amino acid sequence variants of therapeutic proteins can affect their safety and efficacy. Top-down mass spectrometry is well suited for the sequence analysis of intact therapeutic proteins. Fine-tuning of tandem mass spectrometry (MS/ MS) fragmentation conditions is essential for maximizing the amino acid sequence coverage but is often time-consuming. We used topdownr, an automated and integrated multimodal approach to systematically assess high mass accuracy MS/MS fragmentation parameters to characterize filgrastim, a 19 kDa recombinant human granulocyte colony-stimulating factor used in treating neutropenia. A total of 276 different MS/MS conditions were systematically tested, including the following parameters: protein charge state, HCD and CID collision energy, ETD reaction time, ETD supplemental activation, and UVPD activation time. Stringent and accurate evaluation and annotation of the MS/MS data was achieved by requiring a fragment ion mass error of 5 ppm, considering reproducible N-and C-terminal fragment ions only, and excluding internal fragment ion assignments. We report the first EThcD and UVPD MS/MS analysis of intact filgrastim, and these two techniques combined resulted in 98% amino acid sequence coverage. By combining all tested fragmentation modes, we obtained near-complete amino acid sequence coverage (99.4%) of intact filgrastim.

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Section: Resultsmentioning

confidence: 99%

Section: Contribution Of Individual Ms/ms Fragmentation Modes To the ...mentioning

confidence: 92%

Effective Amino Acid Sequencing of Intact Filgrastim by Multimodal Mass Spectrometry with Topdownr

Babović

Shliaha

Gibb

et al. 2022

J. Am. Soc. Mass Spectrom.

View full text Add to dashboard Cite

show abstract

Receptor-binding hydrogen–deuterium exchange mass spectrometry as an additional measurement of biosimilarity

Cited by 1 publication

References 31 publications

Effective Amino Acid Sequencing of Intact Filgrastim by Multimodal Mass Spectrometry with Topdownr

Effective Amino Acid Sequencing of Intact Filgrastim by Multimodal Mass Spectrometry with Topdownr

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