抗体从头测序的组装算法评测及其在泛冠状病毒中和抗体解析中的应用OA

Humanized monoclonal antibodies,generated by grafting complementarity-determining regions(CDRs),offer significant advantages in terms of safety and efficacy as therapeutic agents.The accurate determination of heavy-and light-chain CDR sequences is crucial for the development and production of recombinant antibodies.In this study,we performed de novo sequencing on three humanized pan-coronavirus neutralizing antibodies(3D11,C1520,5A6)utilizing an integrated SP-MEGD pretreatment method combined with a bottom-up mass spectrometry approach.The antibody sequences were reconstructed employing three assembly algorithms:PEAKS AB,Stitch,and Fu-sion.A systematic comparison was conducted to evaluate the performance of these three algorithms,focusing on their assembly coverage and accuracy.The results indicated that the Fusion algorithm achieves 100%coverage and accuracy under the current experimental conditions,for both light and heavy chains across all three antibodies.Furthermore,it maintains stability in high-variability re-gions of CDRs without introducing any additional sequence insertions.In contrast,PEAKS AB and Stitch exhibited fragment missing,insertions,or misassembled segments in certain areas.In conclu-sion,the robust performance demonstrated by the Fusion algorithm provides a reliable foundation for sequence analysis that supports rational design and development efforts concerning pan-coronavirus neutralizing antibodies.