XA-Novo: an accurate and high-throughput mass spectrometry-based de novo sequencing technology for monoclonal antibodies and antibody mixtures
Listed in
This article is not in any list yet, why not save it to one of your lists.Abstract
Elucidating antibody sequences by mass spectrometry (MS)-based de novo sequencing is essential but challenging. Herein, we propose XA-Novo, an accurate, robust, high-throughput de novo sequencing solution that integrates a single-pot multi-enzymatic gradient digestion method with a beam search-based Fusion assembler to accurately decipher complete antibody sequences at both the sample preparation stage and the data analysis phase. We first benchmark XA-Novo using data from multiple known antibodies across various species, demonstrating that it outperforms commercial solutions in identification capacity, data completeness, and antibody reconstruction accuracy. Furthermore, XA-Novo successfully reconstructs three immunotherapeutic antibodies with unknown sequences, and in vitro assays validating that the generated antibodies exhibit equivalent functionality to their commercial counterparts. Furthermore, XA-Novo excels in discriminating mixtures of COVID-19 neutralizing antibodies, achieving over 99% sequence coverage accuracy, further validating the effectiveness and flexibility of our solution. We anticipate that XA-Novo will be a powerful tool for future antibody research applications.
