Glycoengineered recombinant alpha1-antitrypsin results in comparable in vitro and in vivo activities to human plasma-derived protein.
Glycoengineered recombinant alpha1-antitrypsin results in comparable in vitro and in vivo activities to human plasma-derived protein.
复制标题
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
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发表时间：
2024
期刊：
影响因子：
--
通讯作者：
Lewis,NathanE
中科院分区：
文献类型：
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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
α -1抗胰蛋白酶（A1AT）是一种多功能、临床重要、高价值的治疗糖蛋白，可用于治疗α -1抗胰蛋白酶缺乏症、糖尿病、移植物抗宿主病、囊性纤维化和各种病毒感染等多种疾病。目前，fda唯一批准的A1AT疾病治疗方法是人血浆源性A1AT静脉增强治疗。除了供应有限之外，这种方法还存在感染传播的风险，因为它使用的是来自捐赠者的治疗性A1AT。为了解决这些问题，我们试图利用糖工程中国仓鼠卵巢（geCHO-L）细胞产生重组人A1AT (rhA1AT)，该细胞在化学和生物学上与血浆来源的A1AT没有区别。通过删除CHO糖基化机制的9个关键基因，并表达人类ST6GAL1和A1AT基因，我们获得了稳定、高产的geCHO-L细胞系，其产生的rhA1AT具有与血浆源性A1AT （pdA1AT）相同的糖谱。此外，rhA1AT的体外活性和体内半衰期与商用pdA1AT相当。因此，我们预计该平台将有助于生产类似人类的重组血浆蛋白，从而提供更可持续和可靠的治疗来源，具有成本效益，并且在纯度，临床安全性和质量方面得到更好的控制。
Alpha-1-antitrypsin (A1AT) is a multifunctional, clinically important, high value therapeutic glycoprotein that can be used for the treatment of many diseases such as alpha-1-antitrypsin deficiency, diabetes, graft-versus-host-disease, cystic fibrosis and various viral infections. Currently, the only FDA-approved treatment for A1AT disorders is intravenous augmentation therapy with human plasma-derived A1AT. In addition to its limited supply, this approach poses a risk of infection transmission, since it uses therapeutic A1AT harvested from donors. To address these issues, we sought to generate recombinant human A1AT (rhA1AT) that is chemically and biologically indistinguishable from its plasma-derived counterpart using glycoengineered Chinese Hamster Ovary (geCHO-L) cells. By deleting nine key genes that are part of the CHO glycosylation machinery and expressing the human ST6GAL1 and A1AT genes, we obtained stable, high producing geCHO-L lines that produced rhA1AT having an identical glycoprofile to plasma-derived A1AT (pdA1AT). Additionally, the rhA1AT demonstrated in vitro activity and in vivo half-life comparable to commercial pdA1AT. Thus, we anticipate that this platform will help produce human-like recombinant plasma proteins, thereby providing a more sustainable and reliable source of therapeutics that are cost-effective and better-controlled with regard to purity, clinical safety and quality.