ArticleJournal of the American Society for Mass Spectrometry2025
Assessing the Impact of CDR Deamidation and Isomerization on mAb Target Binding Using Affinity-Resolved 2D-SEC-SCX-MS Analysis.
Article in Journal of the American Society for Mass Spectrometry, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
What it found
Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.
The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
0 citing papers in PubMed.
No citing paper in PubMed yet.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Post-translational modifications (PTMs) within the Complementarity-Determining Regions (CDRs) of monoclonal antibodies (mAbs) can impact their target-binding capabilities, making them potential critical quality attributes (CQAs) during therapeutic mAb development. Conventional methods for assessing PTM criticality often involve variant enrichment followed by target-binding testing, which face limitations in throughput and complexity. To address these challenges, affinity enrichment-based strategies have emerged, offering a valuable alternative for PTM assessment. Notably, the combination of competitive binding, size exclusion chromatography (SEC) separation, and MS detection, has proven highly effective in assessing the criticality of PTMs in a multiplexed fashion. Recently, we introduced a new technique termed affinity-resolved SEC-MS, which employs SEC to separate free and target-bound mAbs, followed by postcolumn denaturation (PCD)-assisted intact mass measurements. While highly effective in interrogating PTMs associated with significant mass shifts, this technique is less suitable for studying PTMs with subtle mass changes, such as asparagine (Asn) deamidation and aspartic acid (Asp) isomerization - two prevalent and often critical PTMs found in therapeutic mAbs. To overcome this limitation, we combined affinity-resolved SEC separation with online strong cation exchange chromatography-MS (SCX-MS) analysis. This 2D-LC-MS approach leverages the excellent selectivity of SCX separation for mAb CDR modifications at the Fab fragment level, enabling effective evaluation of Asn deamidation and Asp isomerization with site-specific resolution. The utility of this new approach was demonstrated through two case studies: examining a single Asn deamidation in an in-house mAb and assessing multiple site-specific Asn deamidation and Asp isomerization in trastuzumab, all occurring within the CDRs. Additionally, we detailed a quantitative approach to estimate the relative fold change in dissociation constant (
Indexed as
Identifiers
What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.