“Monoclonal antibodies (mAbs) are a rapidly advancing class of therapeutic glycoproteins that possess wide clinical utility owing to their biocompatibility, high antigen specificity, and targeted immune stimulation. These therapeutic properties depend greatly on the composition of the immunoglobulin G structure, both in terms of primary sequence and post-translational modifications; however, large-scale production in cell culture often results in heterogeneous mixtures that can profoundly affect clinical safety and efficacy. This places a high demand on analytical methods that afford comprehensive structural characterization of mAbs to ensure their stringent quality control.”
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Cotham VC, Brodbelt JS. 2016. Characterization of Therapeutic Monoclonal Antibodies at the Subunit-Level using Middle-Down 193 nm Ultraviolet Photodissociation. Anal. Chem. 88(7):4004–13
Resemann A, Jabs W, Wiechmann A, Wagner E, Colas O, et al. 2016. Full validation of therapeutic antibody sequences by middle-up mass measurements and middle-down protein sequencing. mAbs. 8(2):318–30
Pan J, Zhang S, Chou A, Hardie DB, Borchers CH. 2015. Fast Comparative Structural Characterization of Intact Therapeutic Antibodies Using Hydrogen–Deuterium Exchange and Electron Transfer Dissociation. Anal. Chem. 87(12):5884–90
Dekker L, Wu S, Vanduijn M, Tolić N, Stingl C, et al. 2014. An integrated top-down and bottom-up proteomic approach to characterize the antigen-binding fragment of antibodies. Proteomics. 14(10):1239–48
Zhang H, Cui W, Gross ML. 2014. Mass spectrometry for the biophysical characterization of therapeutic monoclonal antibodies. FEBS Letters. 588(2):308–17
Mao Y, Valeja SG, Rouse JC, Hendrickson CL, Marshall AG. 2013. Top-Down Structural Analysis of an Intact Monoclonal Antibody by Electron Capture Dissociation-Fourier Transform Ion Cyclotron Resonance-Mass Spectrometry. Anal. Chem. 85(9):4239–46
Tsybin YO, Fornelli L, Stoermer C, Luebeck M, Parra J, et al. 2011. Structural Analysis of Intact Monoclonal Antibodies by Electron Transfer Dissociation Mass Spectrometry. Anal. Chem. 83(23):8919–27