Michaela Helble, Jacqueline Chu, Kaitlyn Flowers, Abigail R Trachtman, Alana Huynh, Amber Kim, Nicholas Shupin, Casey E Hojecki, Ebony N Gary, Shahlo Solieva, Elizabeth M Parzych, David B Weiner, Daniel W Kulp, Ami Patel
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引用次数: 0
摘要
单克隆抗体是一类重要的生物制剂,美国食品和药物管理局(FDA)/欧盟共批准了 160 多种单克隆抗体药物。全球获得重组单克隆抗体的一个重要瓶颈是其生产、储存和销售的复杂性。最近,基因编码的方法(如 mRNA、DNA 或病毒递送)越来越受欢迎,但确保抗体在宿主体内的生物相关表达水平仍然是一个关键问题。利用合成 DNA 平台,我们研究了抗体结构和序列对体内表达的作用。SARS-COV2 抗体 2196 最近被设计成一种 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 "的方法,我们通过合理的设计和结构建模研究了 DMAb-2196 可调节体内表达的特征。将这些结果与抗体序列的自然变异进行比较后,我们开发出了一种抗体频率评分,通过利用抗体库数据集来帮助预测可改善表达的突变。我们证明,通过该评分确定的单个氨基酸突变可将体内表达量提高 2 倍,突变组合也可提高表达量。通过这项分析,我们开发出了一种通用的管道,可以释放体内输送治疗性抗体的潜力,适用于多种适应症。
Structure and sequence engineering approaches to improve in vivo expression of nucleic acid-delivered antibodies.
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.
期刊介绍:
Molecular Therapy is the leading journal for research in gene transfer, vector development, stem cell manipulation, and therapeutic interventions. It covers a broad spectrum of topics including genetic and acquired disease correction, vaccine development, pre-clinical validation, safety/efficacy studies, and clinical trials. With a focus on advancing genetics, medicine, and biotechnology, Molecular Therapy publishes peer-reviewed research, reviews, and commentaries to showcase the latest advancements in the field. With an impressive impact factor of 12.4 in 2022, it continues to attract top-tier contributions.