Middle-Down 193-nm Ultraviolet Photodissociation for Unambiguous Antibody Identification and its Implications for Immunoproteomic Analysis

Analytical Chemistry
Victoria C CothamJennifer S Brodbelt

Abstract

Mass spectrometry (MS) has emerged as a powerful tool within the growing field of immunoproteomics, which aims to understand antibody-mediated immunity at the molecular-level based on the direct determination of serological antibody repertoire. To date, these methods have relied on the use of high-resolution bottom-up proteomic strategies that require effective sampling and characterization of low abundance peptides derived from the antigen-binding domains of polyclonal antibody mixtures. Herein, we describe a method that uses restricted Lys-C enzymatic digestion to increase the average mass of proteolytic IgG peptides (≥4.5 kDa) and produce peptides which uniquely derive from single antibody species. This enhances the capacity to discriminate between very similar antibodies present within polyclonal mixtures. Furthermore, our use of 193-nm ultraviolet photodissociation (UVPD) improves spectral coverage of the antibody sequence relative to conventional collision- and electron-based fragmentation methods. We apply these methods to both a monoclonal and an antibody mixture. By identifying from a database search of approximately 15 000 antibody sequences those which compose the mixture, we demonstrate the analytical potential of m...Continue Reading

References

Apr 14, 1983·Nature·S Tonegawa
Oct 13, 2005·Rapid Communications in Mass Spectrometry : RCM·Jeong Hee MoonMyung Soo Kim
Jul 16, 2008·Journal of Proteomics·Xiangming Fang, Wei-Wei Zhang
Dec 17, 2008·Nature Reviews. Immunology·Facundo D Batista, Naomi E Harwood
Feb 26, 2009·Mass Spectrometry Reviews·James P Reilly
Oct 31, 2009·Proceedings of the National Academy of Sciences of the United States of America·Jacob GlanvilleJaume Pons
Mar 5, 2010·The Journal of Allergy and Clinical Immunology·Harry W Schroeder, Lisa Cavacini
Apr 15, 2010·Journal of Proteome Research·Dominique de CostaTheo M Luider
Jun 19, 2010·Journal of Proteome Research·Joe CannonCatherine Fenselau
Jun 29, 2010·Journal of Proteome Research·James A MadsenJennifer S Brodbelt
Jul 10, 2010·The Journal of Biological Chemistry·Martijn M VanDuijnTheo M Luider
Jul 21, 2010·Nature Immunology·Christopher C GoodnowRobert Brink
Nov 26, 2010·Analytical and Bioanalytical Chemistry·Lennard J M DekkerTheo M Luider
Apr 29, 2011·Journal of the European Academy of Dermatology and Venereology : JEADV·G Murphy, K Reich
Aug 11, 2011·PloS One·Ramy ArnaoutSergei B Koralov
Sep 29, 2011·Journal of the American Society for Mass Spectrometry·Lisa A VasicekJennifer S Brodbelt
Mar 27, 2012·Nature Biotechnology·Wan Cheung CheungRoberto D Polakiewicz
Jun 19, 2012·Nature Methods·Cong WuNeil L Kelleher
Sep 26, 2012·Nature Reviews. Immunology·Patrick C Wilson, Sarah F Andrews
Feb 6, 2013·Proceedings of the National Academy of Sciences of the United States of America·Yariv WineGeorge Georgiou
Feb 27, 2013·Chemical Reviews·Yaoyang ZhangJohn R Yates
Mar 14, 2013·Frontiers in Immunology·Bryan S Briney, James E Crowe
Mar 16, 2013·Journal of Mass Spectrometry : JMS·Joe R CannonCatherine Fenselau
May 24, 2013·Journal of the American Chemical Society·Jared B ShawJennifer S Brodbelt
Jan 21, 2014·Nature Biotechnology·George GeorgiouStephen R Quake
Jan 23, 2014·Analytical Chemistry·Joe R CannonJennifer S Brodbelt
Jan 29, 2014·Proceedings of the National Academy of Sciences of the United States of America·Jason J LavinderGeorge Georgiou
Apr 2, 2014·Analytical Chemistry·Daniel R BoutzEdward M Marcotte
Sep 11, 2014·Analytical Chemistry·Kristina SrzentićYury O Tsybin
Dec 17, 2014·Journal of the American Society for Mass Spectrometry·Marion GirodPhilippe Dugourd
Dec 24, 2014·Nature Reviews. Rheumatology·William H Robinson
Feb 24, 2015·Nature Reviews. Immunology·Stephen L NuttLynn M Corcoran
Apr 1, 2015·The FEBS Journal·Liana Tsiatsiani, Albert J R Heck
May 27, 2015·Bioinformatics·Yana SafonovaJennie Lill
Dec 3, 2015·Analytical Chemistry·Jennifer S Brodbelt
Apr 29, 2016·Nature Protocols·Piero GiansantiAlbert J R Heck
Aug 16, 2016·Current Opinion in Structural Biology·Mark L Chiu, Gary L Gilliland

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Citations

Aug 8, 2018·Clinical and Experimental Immunology·J J WangT P Gordon
Apr 24, 2019·Nature Methods·Pascal EgloffMarkus A Seeger
Apr 28, 2020·Journal of Proteome Research·Jianhui ChengDavid D Y Chen
Dec 19, 2019·Chemical Reviews·Jennifer S BrodbeltInês Santos
Nov 2, 2019·Analytical Chemistry·Luis A MaciasJennifer S Brodbelt

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