Direct Mass Spectrometry-Based Detection and Antibody Sequencing of Monoclonal Gammopathy of Undetermined Significance from Patient Serum: A Case Study
peer-reviewed · Journal of Proteome Research · 2023
| Date | 2023-09-01 |
| Type | peer-reviewed |
| Venue | Journal of Proteome Research |
| Publisher | American Chemical Society (ACS) |
| Contribution | downstream-application |
| DOI | 10.1021/acs.jproteome.3c00330 |
| Citations (OpenAlex) | 30 |
| Venue 2-year citedness | 3.83 |
Abstract
Monoclonal gammopathy of undetermined significance (MGUS) is a plasma cell disorder characterized by the presence of a predominant monoclonal antibody (i.e., M-protein) in serum, without clinical symptoms. Here we present a case study in which we detect MGUS by liquid-chromatography coupled with mass spectrometry (LC-MS) profiling of IgG1 in human serum. We detected a Fab-glycosylated M-protein and determined the full heavy and light chain sequences by bottom-up proteomics techniques using multiple proteases, further validated by top-down LC-MS. Moreover, the composition and location of the Fab-glycan could be determined in CDR1 of the heavy chain. The outlined approach adds to an expanding mass spectrometry-based toolkit to characterize monoclonal gammopathies such as MGUS and multiple myeloma, with fine molecular detail. The ability to detect monoclonal gammopathies and determine M-protein sequences straight from blood samples by mass spectrometry provides new opportunities to understand the molecular mechanisms of such diseases.
Methods and tools
- MGUS M-protein sequencing from serum: Detected a monoclonal gammopathy in patient serum and sequenced the full Fab-glycosylated M-protein by multi-protease bottom-up de novo sequencing with template assembly, validated by top-down MS.
Methods it uses
- PEAKS: Commercial DP-based de novo
- Stitch: Assembles de novo peptides from Casanovo, PEAKS, pNovo and MaxNovo into full antibody sequences, and corrects the two error classes that assembly alone cannot: mass coincidences, where a different residue combination matches the same mass, and I/L ambiguity.
Data deposited
- Direct mass spectrometry-based detection and antibody sequencing of Monoclonal Gammopathy of Undetermined Significance from patient serum — as deposited · PXD042266
Cites (12)
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