A novel C-terminal modification method enhanced the yield of human papillomavirus L1 or chimeric L1-L2 virus-like particles in the baculovirus system.
A novel C-terminal modification method enhanced the yield of human papillomavirus L1 or chimeric L1-L2 virus-like particles in the baculovirus system.
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DOI：
10.3389/fbioe.2022.1073892
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发表时间：
2022
影响因子：
5.7
通讯作者：
Xu, Xuemei
中科院分区：
文献类型：
--
作者：
Ma, Mingrao;Xia, Baicheng;Wang, Zhirong;Hao, Yaru;Zhang, Ting;Xu, Xuemei
杆状病毒系统生产的人乳头瘤病毒（HPV）主要衣壳蛋白L1病毒样颗粒（VLPs）具有良好的安全性和免疫原性，但相对较高的生产成本是广泛商业化的实质性障碍，特别是在生产多价疫苗方面。本研究开发了一种新的方法，即c端碱性氨基酸（aa）取代，以提高该体系中VLP和嵌合VLP （cVLP）的产量。构建了5种HPV类型的一系列突变体，包括3个L1型VLPs （6L1、11L1和52L1）和2个L1- l2型cVLPs （16L1-33L2、58L1-16L2）。我们发现，大多数突变体在Sf9细胞中都表现出较高的蛋白表达，其中优势突变体6L1CS4、11L1CS3、52L1m4∆N13CS1、16L1-33L2 CS1和58L1-16L2 CS3的产量分别达到40、35、20、35和60 mg/L，分别提高了4.2倍、7.3倍、5倍、2.5倍和3.4倍，且具有较强的免疫原性和较好的稳定性。此外，我们发现稳态mRNA水平的增加可能在促进L1蛋白表达中起关键作用。我们的研究结果表明，这种新方法具有成本效益，可用于降低L1- l2 VLPs和L1- l2 cVLPs的生产成本，以开发广泛保护和负担得起的多价HPV疫苗。
Human papillomavirus (HPV) major capsid protein L1 virus-like particles (VLPs) produced in the baculovirus system showed excellent safety and immunogenicity, but the relatively high production cost stands as a substantial barrier to extensive commercialization, especially in producing multivalent vaccines. Here, a novel method, C-terminal basic amino acid (aa) substitution, was developed for increasing VLP and chimeric VLP (cVLP) production in this system. A series of mutants of five HPV types, including three L1 VLPs (6L1, 11L1, and 52L1) and two L1-L2 cVLPs (16L1-33L2, 58L1-16L2), were constructed. We found that most mutants exhibited higher protein expression in Sf9 cells, among which the yields of the superior mutants, 6L1CS4, 11L1CS3, 52L1m4∆N13CS1, 16L1-33L2 CS1, and 58L1-16L2 CS3, were up to 40, 35, 20, 35, and 60 mg/L, which respectively increased by 4.2-, 7.3-, 5-, 2.5-, and 3.4-fold, and they also showed robust immunogenicity and great stabilities. Additionally, we found that the increased level of steady-state mRNA may play a crucial role in promoting L1 protein expression. Our results demonstrated that this novel method was cost-effective and can be used to reduce the production costs of L1 VLPs and L1-L2 cVLPs to develop broadly protective and affordable multivalent HPV vaccines.
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影响因子：
16.6
作者：
Krarup A;Truan D;Furmanova-Hollenstein P;Bogaert L;Bouchier P;Bisschop IJM;Widjojoatmodjo MN;Zahn R;Schuitemaker H;McLellan JS;Langedijk JPM
通讯作者：
Langedijk JPM
影响因子：
3.7
作者：
Huber B;Schellenbacher C;Jindra C;Fink D;Shafti-Keramat S;Kirnbauer R
通讯作者：
Kirnbauer R
影响因子：
6.4
作者：
Kann, Hanna;Lehtinen, Matti;Faust, Helena
通讯作者：
Faust, Helena
影响因子：
5.4
作者：
Boxus, Mathieu;Fochesato, Michel;Giannini, Sandra L.
通讯作者：
Giannini, Sandra L.
影响因子：
3.5
作者：
Neeper, MP;Hofmann, KJ;Jansen, KU
通讯作者：
Jansen, KU