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J Am Soc Mass Spectrom. 2017 May;28(5):803-810. doi: 10.1007/s13361-016-1580-0. Epub 2017 Jan 19.

Automated Antibody De Novo Sequencing and Its Utility in Biopharmaceutical Discovery.

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Protein Metrics Inc, 1622 San Carlos Ave, Suite C, San Carlos, CA, 94070, USA.
Protein Metrics Inc, 1622 San Carlos Ave, Suite C, San Carlos, CA, 94070, USA.
Langone Medical Center, New York University, 430 East 29th street, 8th floor room 860, New York, NY, 10016, USA.
Janssen Research and Development, LLC, 1400 McKean Road, Spring House, PA, 19477, USA.


Applications of antibody de novo sequencing in the biopharmaceutical industry range from the discovery of new antibody drug candidates to identifying reagents for research and determining the primary structure of innovator products for biosimilar development. When murine, phage display, or patient-derived monoclonal antibodies against a target of interest are available, but the cDNA or the original cell line is not, de novo protein sequencing is required to humanize and recombinantly express these antibodies, followed by in vitro and in vivo testing for functional validation. Availability of fully automated software tools for monoclonal antibody de novo sequencing enables efficient and routine analysis. Here, we present a novel method to automatically de novo sequence antibodies using mass spectrometry and the Supernovo software. The robustness of the algorithm is demonstrated through a series of stress tests. Graphical Abstract ᅟ.


Antibody; Byonic; De novo sequencing; Mass spectrometry; Supernovo; mAb

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