De Novo MS/MS Sequencing of Native Human Antibodies
peer-reviewed · Journal of Proteome Research · 2017
| Date | 2017-01-06 |
| Type | peer-reviewed |
| Venue | Journal of Proteome Research |
| Publisher | American Chemical Society (ACS) |
| Contribution | downstream-application |
| DOI | 10.1021/acs.jproteome.6b00608 |
| Citations (OpenAlex) | 68 |
| Venue 2-year citedness | 3.48 |
Abstract
One direct route for the discovery of therapeutic human monoclonal antibodies (mAbs) involves the isolation of peripheral B cells from survivors/sero-positive individuals after exposure to an infectious reagent or disease etiology, followed by single-cell sequencing or hybridoma generation. Peripheral B cells, however, are not always easy to obtain and represent only a small percentage of the total B-cell population across all bodily tissues. Although it has been demonstrated that tandem mass spectrometry (MS/MS) techniques can interrogate the full polyclonal antibody (pAb) response to an antigen in vivo, all current approaches identify MS/MS spectra against databases derived from genetic sequencing of B cells from the same patient. In this proof-of-concept study, we demonstrate the feasibility of a novel MS/MS antibody discovery approach in which only serum antibodies are required without the need for sequencing of genetic material. Peripheral pAbs from a cytomegalovirus-exposed individual were purified by glycoprotein B antigen affinity and de novo sequenced from MS/MS data. Purely MS-derived mAbs were then manufactured in mammalian cells to validate potency via antigen-binding ELISA. Interestingly, we found that these mAbs accounted for 1 to 2% of total donor IgG but were not detected in parallel sequencing of memory B cells from the same patient.
Methods and tools
- Native antibody de novo MS/MS: De novo MS/MS sequencing workflow for native human antibodies.
Cites (5)
- Sequencing-Grade De novo Analysis of MS/MS Triplets (CID/HCD/ETD) From Overlapping Peptides (2013) both
- Shotgun Protein Sequencing with Meta-contig Assembly (2012) both
- Automated de novo protein sequencing of monoclonal antibodies (2008) both
- Shotgun Protein Sequencing by Tandem Mass Spectra Assembly (2004) semanticscholar
- De novo peptide sequencing via tandem mass spectrometry (1999) both
Cited by (10)
- XA-Novo: an accurate and high-throughput mass spectrometry-based de novo sequencing technology for monoclonal antibodies and antibody mixtures (2026) both
- Simultaneous polyclonal antibody sequencing and epitope mapping by cryo electron microscopy and mass spectrometry (2025) crossref
- Multienzyme deep learning models improve peptide de novo sequencing by mass spectrometry proteomics (2023) both
- Uncovering Hidden Members and Functions of the Soil Microbiome Using De Novo Metaproteomics (2022) semanticscholar
- Human plasma IgG1 repertoires are simple, unique, and dynamic (2021) both
- Highly Robust de Novo Full-Length Protein Sequencing (2021) both
- pNovo 3: precise de novo peptide sequencing using a learning-to-rank framework (2019) both
- Postnovo: Postprocessing Enables Accurate and FDR-Controlled de Novo Peptide Sequencing (2018) crossref
- A potential golden age to come—current tools, recent use cases, and future avenues for de novo sequencing in proteomics (2018) crossref
- Evaluating de novo sequencing in proteomics: already an accurate alternative to database-driven peptide identification? (2017) crossref