Introduction to De Novo Protein Sequencing

Abstract Monoclonal antibodies (mAbs) are on the rise as biologics in the biopharma industry. To develop robut mAb biologics, it is vital to fully characterize the protein, including its primary sequence, mutations, and important post-translational modifications. Conventionally, this data is gathered by performing several distinct biochemical and molecular biology experiments, which can be time-consuming and costly. Furthermore, when proteomics-based mAb characterization is undertaken, many laboratories lack the bioinformatics capacity and still rely on human analysis, which can result in errors, or in homology-based databases which can introduce bias. As such, traditional antibody characterization may be lengthy and error-prone. Here, we automated a protein sequencing workflow, and show that a fully automated LC-MS/MS-based protein sequencing data analysis can efficiently and fully decode the primary amino acid sequence of an antibody, in addition to denoting post-translational modification (PTM) sites critical for antibody characterization required in biologics development.

Key Takeaways De novo protein sequencing is capable of fully sequencing an antibody De novo protein sequencing can pinpoint PTM sites and the type of PTM De novo protein sequencing is useful tool for antibody characterization

Introduction to De Novo Protein Sequencing

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