Monoclonal antibodies are an important class of biologics with over 160 Food and Drug Administration/European Union-approved drugs. A significant bottleneck to global accessibility of recombinant monoclonal antibodies stems from complexities related to their production, storage, and distribution. Recently, gene-encoded approaches such as mRNA, DNA, or viral delivery have gained popularity, but ensuring biologically relevant levels of antibody expression in the host remains a critical issue. Using a synthetic DNA platform, we investigated the role of antibody structure and sequence toward in vivo expression. SARS-CoV-2 antibody 2196 was recently engineered as a DNA-encoded monoclonal antibody (DMAb-2196). Utilizing an immunoglobulin heavy and light chain “chain-swap” methodology, we interrogated features of DMAb-2196 that can modulate in vivo expression through rational design and structural modeling. Comparing these results to natural variation of antibody sequences resulted in development of an antibody frequency score that aids in the prediction of expression-improving mutations by leveraging antibody repertoire datasets. We demonstrate that a single amino acid mutation identified through this score increases in vivo expression up to 2-fold and that combinations of mutations can also enhance expression. This analysis has led to a generalized pipeline that can unlock the potential for in vivo delivery of therapeutic antibodies across many indications.

中文翻译：

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提高核酸递送抗体体内表达的结构和序列工程方法


单克隆抗体是一类重要的生物制剂，有 160 多种药物获得美国食品药品监督管理局/欧盟批准。重组单克隆抗体全球可及性的一个重大瓶颈源于与其生产、储存和分销相关的复杂性。最近，mRNA、DNA 或病毒递送等基因编码方法越来越受欢迎，但确保宿主中抗体表达的生物学相关水平仍然是一个关键问题。使用合成 DNA 平台，我们研究了抗体结构和序列对 体内表达的作用。SARS-CoV-2 抗体 2196 最近被设计为 DNA 编码的单克隆抗体 （DMAb-2196）。利用免疫球蛋白重链和轻链“换链”方法，我们询问了 DMAb-2196 的特征，这些特征可以通过合理的设计和结构建模来 调节体内表达。将这些结果与抗体序列的自然变异进行比较，得出了抗体频率评分，该评分有助于通过利用抗体库数据集来预测改善表达的突变。我们证明，通过该评分鉴定的单个氨基酸突变可将体内表达增加 多达 2 倍，并且突变组合也可以增强表达。该分析导致了一个通用的管道，可以释放在许多适应症中体内递送治疗性抗体的潜力 。