Novel constructs and 1-step chromatography protocols for the production of Porcine Circovirus 2d (PCV2d) and Circovirus 3 (PCV3) subunit vaccine candidates
Novel constructs and 1-step chromatography protocols for the production of Porcine Circovirus 2d (PCV2d) and Circovirus 3 (PCV3) subunit vaccine candidates
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DOI：
10.1016/j.fbp.2021.10.001
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发表时间：
2021-11-25
影响因子：
4.6
通讯作者：
Robinson, Colin
中科院分区：
文献类型：
--
作者：
Peswani, Amber R.;Narkpuk, Jaraspim;Robinson, Colin
猪圆环病毒2型（PCV 2）是全球养猪业的主要问题。虽然大多数市售疫苗是基于原始PCV 2a基因型，但目前的主要基因型是PCV 2d。基因型间的显著差异可能导致交叉保护不完全。此外，目前大多数亚单位PCV 2疫苗是由昂贵的昆虫细胞培养技术产生的。在这项工作中，我们提出了一种新的工作流程，用于生产更新且相对便宜的PCV 2d候选疫苗。在大肠杆菌补料分批发酵系统中表达后，采用简单的一步离子交换层析纯化方案，纯化的PCV 2d抗原产量达到1g/L细菌培养物以上。使用类似的方法，我们还证明了嵌合PCV 2d-PCV 3衣壳构建体的基于PCV 2d的抗原产量甚至更高，所述嵌合PCV 2d-PCV 3衣壳构建体在发酵期间被切割以释放PCV 2d和PCV 3相关多肽。尽管通过分子排阻色谱分析，来自该方案的基于PCV 2d的重组蛋白未形成病毒样颗粒，但其可在免疫动物中有效诱导captain特异性抗体和PCV 2d中和抗体，表明其具有作为新候选疫苗的显著潜力，可轻松以商业规模生产。(c)2021年由Elsevier B. V.代表化学工程师学会出版。
Porcine circovirus type 2 (PCV2) has been a major problem for the pig production industry worldwide for decades. While the majority of commercially available vaccines are based on the original PCV2a genotype, the current dominant genotype is PCV2d. The notable differ-ences between genotypes could lead to incomplete cross-protection. Moreover, most current subunit PCV2 vaccines are generated from expensive insect cell culture technology. In this work, we present a new workflow for production of an updated and relatively inexpensive PCV2d vaccine candidate. After expression in fed-batch Escherichia coli fermentation sys-tems with a simple one-step ion-exchange chromatography purification protocol, the yield of purified PCV2d-based antigen reached over 1 g per litre bacterial culture. Using similar procedures, we also demonstrated even higher PCV2d-based antigen yields from a chimeric PCV2d-PCV3 capsid construct, which is cleaved during fermentation to release PCV2d-and PCV3-related polypeptides. Although the PCV2d-based recombinant protein from this pro-tocol did not form viral-like particles as analysed by size-exclusion chromatography, it could effectively induce capsid-specific and PCV2d-neutralising antibodies in immunised animals, indicating significant potential as a new vaccine candidate that can be easily manufactured at commercial scale. (c) 2021 Published by Elsevier B.V. on behalf of Institution of Chemical Engineers.