Reverse-engineering the anti-MUC1 antibody 139H2 by mass spectrometry–based de novo sequencing
peer-reviewed · Life Science Alliance · 2024
| Date | 2024-06-01 |
| Type | peer-reviewed |
| Venue | Life Science Alliance |
| Publisher | Life Science Alliance, LLC |
| Contribution | downstream-application |
| DOI | 10.26508/lsa.202302366 |
| Citations (OpenAlex) | 11 |
Preprint version: Reverse engineering the anti-MUC1 hybridoma antibody 139H2 by mass spectrometry-based de novo sequencing (2023-07-05, bioRxiv)
Abstract
Mucin 1 (MUC1) is a transmembrane mucin expressed at the apical surface of epithelial cells at mucosal surfaces. MUC1 has a barrier function against bacterial invasion and is well known for its aberrant expression and glycosylation in adenocarcinomas. The MUC1 extracellular domain contains a variable number of tandem repeats (VNTR) of 20 amino acids, which are heavily O -linked glycosylated. Monoclonal antibodies against the MUC1 VNTR are powerful research tools with applications in the diagnosis and treatment of MUC1-expressing cancers. Here, we report direct mass spectrometry-based sequencing of anti-MUC1 hybridoma-derived 139H2 IgG, enabling reverse-engineering of the functional recombinant monoclonal antibody. The crystal structure of the 139H2 Fab fragment in complex with the MUC1 epitope was solved, revealing the molecular basis of 139H2 binding specificity to MUC1 and its tolerance to O -glycosylation of the VNTR. The available sequence of 139H2 will allow further development of MUC1-related diagnostic, targeting, and treatment strategies.
Methods and tools
- Anti-MUC1 139H2 reverse engineering: Reverse-engineers the anti-MUC1 antibody 139H2 by mass spectrometry-based de novo sequencing, MUC1 being a mucin whose aberrant expression and glycosylation matter in adenocarcinoma.
Data deposited
- Sequencing the anti-MUC1 hybridoma antibody 139H2 (as deposited) · PXD043489
Cites (7)
- Into the Dark Serum Proteome: Personalized Features of IgG1 and IgA1 Repertoires in Severe COVID-19 Patients (2024) both
- Direct Mass Spectrometry-Based Detection and Antibody Sequencing of Monoclonal Gammopathy of Undetermined Significance from Patient Serum: A Case Study (2023) both
- Template-Based Assembly of Proteomic Short Reads For De Novo Antibody Sequencing and Repertoire Profiling (2022) both
- Human plasma IgG1 repertoires are simple, unique, and dynamic (2021) both
- Mass Spectrometry-Based De Novo Sequencing of Monoclonal Antibodies Using Multiple Proteases and a Dual Fragmentation Scheme (2021) both
- Deep learning enables de novo peptide sequencing from data-independent-acquisition mass spectrometry (2018) both
- De novo peptide sequencing by deep learning (2017) both
Cited by (5)
- XA-Novo: an accurate and high-throughput mass spectrometry-based de novo sequencing technology for monoclonal antibodies and antibody mixtures (2026) both
- Structural Basis for Postfusion-Specific Binding to the Respiratory Syncytial Virus F Protein by the Canonical Antigenic Site I Antibody 131–2a (2025) both
- Do-It-Yourself De Novo Antibody Sequencing Workflow that Achieves Complete Accuracy of the Variable Regions (2025) crossref
- Deep Learning Methods for De Novo Peptide Sequencing (2024) crossref
- A Handle on Mass Coincidence Errors in De Novo Sequencing of Antibodies by Bottom-up Proteomics (2024) both