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糖基工程毕赤酵母制备的SARS-CoV-2 S蛋白RBD受体结合区亚单位疫苗可激发中和抗体反应
Bo Liu, Ying Yin, Yuxiao Liu, Tiantian Wang, Peng Sun, Yangqin Ou, Xin Gong, Xuchen Hou, Jun Zhang, Hongguang Ren, Shiqiang Luo, Qian Ke, Yongming Yao, Junjie Xu, Jun Wu
工程（英文） ›› 2022, Vol. 13 ›› Issue (6) : 107-115.
PDF(2953 KB)
PDF(2953 KB)
糖基工程毕赤酵母制备的SARS-CoV-2 S蛋白RBD受体结合区亚单位疫苗可激发中和抗体反应
A Vaccine Based on the Receptor-Binding Domain of the Spike Protein Expressed in Glycoengineered Pichia pastoris Targeting SARS-CoV-2 Stimulates Neutralizing and Protective Antibody Responses
2020年至今，新型冠状病毒SARS-CoV-2已经在全球大流行。新冠疫苗被寄于厚望用以减轻防控压力，成为解决疫情危机的有效手段之一。SARS-CoV-2通过S蛋白受体结合区RBD与宿主细胞ACE2受体结合侵染机体。本文提供了一种利用新型糖基工程毕赤酵母表达系统，制备基于S蛋白RBD重组亚单位候选疫苗的方法。该糖基工程酵母的糖基化修饰途径经过人源化改造后，具有类似于哺乳动物细胞糖基化修饰特点。研究发现，RBD候选疫苗可以有效地诱导小鼠产生高滴度的抗RBD特异性抗体，含Al(OH)3和CpG双佐剂的RBD免疫组小鼠产生的特异性抗体滴度和病毒中和抗体滴度显著地高于Al(OH)3单佐剂组，且中和抗体可以在小鼠体内持续6个月以上。综上所述，本文利用糖基工程酵母制备了SARS-CoV-2 S蛋白RBD糖蛋白疫苗，该疫苗能够诱导小鼠产生高滴度中和抗体，同时提供了一种可制备具有无岩藻糖的复杂型N-糖基化修饰重组蛋白的方法。
In 2020 and 2021, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a novel coronavirus, caused a global pandemic. Vaccines are expected to reduce the pressure of prevention and control, and have become the most effective strategy to solve the pandemic crisis. SARS-CoV-2 infects the host by binding to the cellular receptor angiotensin converting enzyme 2 (ACE2) via the receptor-binding domain (RBD) of the surface spike (S) glycoprotein. In this study, a candidate vaccine based on a RBD recombinant subunit was prepared by means of a novel glycoengineered yeast Pichia pastoris expression system with characteristics of glycosylation modification similar to those of mammalian cells. The candidate vaccine effectively stimulated mice to produce high-titer anti-RBD specific antibody. Furthermore, the specific antibody titer and virus-neutralizing antibody (NAb) titer induced by the vaccine were increased significantly by the combination of the double adjuvants Al(OH)3 and CpG. Our results showed that the virus-NAb lasted for more than six months in mice. To summarize, we have obtained a SARS-CoV-2 vaccine based on the RBD of the S glycoprotein expressed in glycoengineered Pichia pastoris, which stimulates neutralizing and protective antibody responses. A technical route for fucose-free complex-type N-glycosylation modified recombinant subunit vaccine preparation has been established.
冠状病毒 / SARS-CoV-2 / 疫苗 / 酵母 / 受体结合区（RBD）
Coronavirus / SARS-CoV-2 / Vaccine / Yeast / Receptor-binding domain (RBD)
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