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. 2017 May;28(5):827-838.
doi: 10.1007/s13361-017-1602-6. Epub 2017 Feb 28.

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

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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

Lidong He et al. J Am Soc Mass Spectrom. .

Erratum in

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.

Keywords: CID; Collision-induced dissociation; ETD; Electron transfer dissociation; Electrospray; FTMS; Fourier transform; Ion cyclotron resonance; Isotype; MS/MS; Middle-down; Multiple myeloma; Variable region.

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References

    1. Clin Vaccine Immunol. 2006 Feb;13(2):277-80 - PubMed
    1. Mass Spectrom Rev. 2009 Jan-Feb;28(1):147-76 - PubMed
    1. J Neuroimmunol. 2015 Nov 15;288:123-6 - PubMed
    1. Crit Rev Clin Lab Sci. 2013 Jul-Oct;50(4-5):91-102 - PubMed
    1. Clin Cancer Res. 2013 Mar 1;19(5):985-94 - PubMed
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