Glycoengineering of CHO cells to express recombinant alpha-1 antitrypsin

CHO细胞的糖工程表达重组α-1抗胰蛋白酶

• 批准号: 10484110
• 负责人: Nathan Enoch Lewis
• 金额: $ 27.44万
• 依托单位: NEUIMMUNE BIOLOGICS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-22 至 2024-09-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10484110
• 关键词: Animals; Biological Response Modifier Therapy; Blood Circulation; Blood-Borne Pathogens; Cell Line; Cells; Chinese Hamster Ovary Cell; Chronic Obstructive Pulmonary Disease; Clinical; Clinical Research; Clinical Trials; Complex; Contracts; Development; Disease; Drug Kinetics; Engineering; Ensure; Frequencies; Genes; Genetic; Glycoengineering; Glycoproteins; Half-Life; Human; INS gene; In Vitro; Incidence; Knock-out; Lead; Liver diseases; Lung Transplantation; Modification; Mutation; Patient Care; Performance; Pharmaceutical Preparations; Plasma; Post-Translational Protein Processing; Preparation; Prions; Production; Protein Engineering; Protein Isoforms; Proteins; Proteolysis; Pulmonary Emphysema; Rattus; Recombinant Proteins; Recombinants; Replacement Therapy; Risk; Safety; Source; System; Therapeutic; Transfusion; Variant; alpha 1-Antitrypsin; alpha 1-Antitrypsin Deficiency; cost; glycosylation; improved; in vitro activity; in vivo; liver transplantation; oxidation; preclinical study; stem; therapeutic candidate

ABSTRACT Alpha-1 antitrypsin (A1AT) is well-established as a biotherapeutic used for the treatment of alpha-1 antitrypsin deficiency, and shows promise for treating a variety of other diseases. However, A1AT augmentation therapy carries unnecessary risk since it relies upon the weekly transfusion of plasma-derived product, which presents supply chain and contaminant risks. This could be remedied with recombinant A1AT, if it adequately matches or improves upon the plasma-derived product, including its post-translational modifications and functional attributes. However, major hurdles to the development of recombinant A1AT exist, since post- translational modifications, such as glycosylation, impact drug activities and half-life, and it has remained challenging to match glycosylation of recombinant A1AT to the plasma derived isoforms. Here we developed a large panel of diverse GMP-ready glycoengineered CHO (geCHO) cell lines that allowed us to discover a host cell line, geCHO-L, that matches glycosylation of the plasma derived product. Here we will demonstrate our platform not only allows us to identify a CHO clone matches the glycosylation of the approved product, but that we can also match the function and half-life of the approved A1AT to enable the manufacturing of a recombinant A1AT to increase the safety and protect the supply of therapeutic A1AT. We further engineer the recombinant A1AT to obtain a product with improved activity and half-life as a candidate therapeutic for alpha-1 antitrypsin deficiency and other diseases.

抽象的 Alpha-1 抗胰蛋白酶 (A1AT) 是公认的用于治疗 alpha-1 的生物治疗药物 抗胰蛋白酶缺乏症，并显示出治疗多种其他疾病的前景。然而，A1AT增强 治疗带来不必要的风险，因为它依赖于每周输注血浆衍生产品，这 存在供应链和污染风险。如果重组 A1AT 能够充分解决这一问题， 匹配或改进血浆衍生产品，包括其翻译后修饰和 功能属性。然而，重组 A1AT 的开发存在主要障碍，因为后 翻译修饰，例如糖基化，影响药物活性和半衰期，并且它仍然存在 将重组 A1AT 的糖基化与血浆来源的异构体相匹配具有挑战性。在这里我们开发了一个 大量不同的 GMP 就绪糖工程 CHO (geCHO) 细胞系使我们能够发现宿主 细胞系 geCHO-L，与血浆衍生产物的糖基化相匹配。在这里我们将展示我们的 平台不仅使我们能够识别与批准产品的糖基化相匹配的 CHO 克隆，而且 我们还可以匹配已批准的 A1AT 的功能和半衰期，以实现重组的生产 A1AT 增加安全性并保护治疗性 A1AT 的供应。我们进一步设计重组 A1AT 获得具有改善的活性和半衰期的产品作为 α-1 抗胰蛋白酶的候选治疗剂 缺乏和其他疾病。

项目成果

Glycoengineered recombinant alpha1-antitrypsin results in comparable in vitro and in vivo activities to human plasma-derived protein.

糖基工程重组 α1-抗胰蛋白酶的体外和体内活性与人血浆衍生蛋白相当。

• DOI: 10.1101/2024.03.27.587088
• 发表时间: 2024
• 期刊: bioRxiv : the preprint server for biology
• 作者: Rocamora,Frances;Schoffelen,Sanne;Arnsdorf,Johnny;Toth,EricA;Abdul,Yunus;Cleveland4th,ThomasE;Bjørn,SaraPetersen;Wu,MinaYingMin;McElvaney,NoelG;Voldborg,BjørnGunnarRude;Fuerst,ThomasR;Lewis,NathanE
• 通讯作者: Lewis,NathanE