Back to Search
Start Over
Analysis of Monoclonal Antibodies in Human Serum as a Model for Clinical Monoclonal Gammopathy by Use of 21 Tesla FT-ICR Top-Down and Middle-Down MS/MS.
- Source :
-
Journal of the American Society for Mass Spectrometry [J Am Soc Mass Spectrom] 2017 May; Vol. 28 (5), pp. 827-838. Date of Electronic Publication: 2017 Feb 28. - Publication Year :
- 2017
-
Abstract
- With the rapid growth of therapeutic monoclonal antibodies (mAbs), stringent quality control is needed to ensure clinical safety and efficacy. Monoclonal antibody primary sequence and post-translational modifications (PTM) are conventionally analyzed with labor-intensive, bottom-up tandem mass spectrometry (MS/MS), which is limited by incomplete peptide sequence coverage and introduction of artifacts during the lengthy analysis procedure. Here, we describe top-down and middle-down approaches with the advantages of fast sample preparation with minimal artifacts, ultrahigh mass accuracy, and extensive residue cleavages by use of 21 tesla FT-ICR MS/MS. The ultrahigh mass accuracy yields an RMS error of 0.2-0.4 ppm for antibody light chain, heavy chain, heavy chain Fc/2, and Fd subunits. The corresponding sequence coverages are 81%, 38%, 72%, and 65% with MS/MS RMS error ~4 ppm. Extension to a monoclonal antibody in human serum as a monoclonal gammopathy model yielded 53% sequence coverage from two nano-LC MS/MS runs. A blind analysis of five therapeutic monoclonal antibodies at clinically relevant concentrations in human serum resulted in correct identification of all five antibodies. Nano-LC 21 T FT-ICR MS/MS provides nonpareil mass resolution, mass accuracy, and sequence coverage for mAbs, and sets a benchmark for MS/MS analysis of multiple mAbs in serum. This is the first time that extensive cleavages for both variable and constant regions have been achieved for mAbs in a human serum background. Graphical Abstract ᅟ.
- Subjects :
- Adalimumab analysis
Amino Acid Sequence
Antibodies, Monoclonal analysis
Fourier Analysis
Humans
Protein Processing, Post-Translational
Software
Spectrometry, Mass, Electrospray Ionization methods
Adalimumab blood
Antibodies, Monoclonal blood
Paraproteinemias blood
Tandem Mass Spectrometry methods
Subjects
Details
- Language :
- English
- ISSN :
- 1879-1123
- Volume :
- 28
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Journal of the American Society for Mass Spectrometry
- Publication Type :
- Academic Journal
- Accession number :
- 28247297
- Full Text :
- https://doi.org/10.1007/s13361-017-1602-6