Biel Thomas G, Faison Talia, Matthews Alicia M, Ortega-Rodriguez Uriel, Falkowski Vincent M, Meek Edward, Bush Xin, Flores Matthew, Johnson Sarah, Wu Wells W, Lehtimaki Mari, Shen Rong-Fong, Agarabi Cyrus, Rao V Ashutosh, Chambers Janice E, Ju Tongzhong
Office of Biotechnology Products, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, Food and Drug Administration Silver Spring Maryland USA.
Department of Comparative Biomedical Sciences, Center for Environmental Health Sciences College of Veterinary Medicine, Mississippi State University Mississippi State Mississippi USA.
Bioeng Transl Med. 2024 Apr 25;9(5):e10666. doi: 10.1002/btm2.10666. eCollection 2024 Sep.
Organophosphate (OP) toxicants remain an active threat to public health and to warfighters in the military. Current countermeasures require near immediate administration following OP exposure and are reported to have controversial efficacies. Acetylcholinesterase (AChE) fused to the human immunoglobulin 1 (IgG1) Fc domain (AChE-Fc) is a potential bioscavenger for OP toxicants, but a reproducible AChE-Fc biomanufacturing strategy remains elusive. This report is the first to establish a comprehensive laboratory-scale bioprocessing strategy that can reproducibly produce AChE-Fc and AChE(W86A)-Fc which is a mutated AChE protein with reduced enzymatic activity. Characterization studies revealed that AChE-Fc and AChE(W86A)-Fc are -glycosylated dimeric fusion glycoproteins but only AChE-Fc had the capability to bind to paraoxon (a model OP). This AChE-Fc fusion glycoprotein bioprocessing strategy can be leveraged during industrial biomanufacturing development, while the research-grade AChE-Fc proteins can be used to determine the potential clinical relevance of the countermeasure against OP toxicants.
有机磷酸酯（OP）毒物仍然对公众健康以及军队中的作战人员构成切实威胁。当前的应对措施要求在接触OP后尽快给药，且据报道其疗效存在争议。与人免疫球蛋白1（IgG1）Fc结构域融合的乙酰胆碱酯酶（AChE）（AChE-Fc）是一种潜在的OP毒物生物清除剂，但可重复的AChE-Fc生物制造策略仍然难以捉摸。本报告首次建立了一种全面的实验室规模生物加工策略，该策略能够可重复地生产AChE-Fc和AChE（W86A）-Fc，后者是一种酶活性降低的突变AChE蛋白。表征研究表明，AChE-Fc和AChE（W86A）-Fc是N-糖基化的二聚体融合糖蛋白，但只有AChE-Fc具有与对氧磷（一种OP模型）结合的能力。这种AChE-Fc融合糖蛋白生物加工策略可在工业生物制造开发过程中加以利用，而研究级别的AChE-Fc蛋白可用于确定针对OP毒物的对策的潜在临床相关性。
