Production of full-length soluble Plasmodium falciparum RH5 protein vaccine using a Drosophila melanogaster Schneider 2 stable cell line system.
Production of full-length soluble Plasmodium falciparum RH5 protein vaccine using a Drosophila melanogaster Schneider 2 stable cell line system.
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DOI：
10.1038/srep30357
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发表时间：
2016-07-26
影响因子：
4.6
通讯作者：
Draper SJ
中科院分区：
文献类型：
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作者：
Hjerrild KA;Jin J;Wright KE;Brown RE;Marshall JM;Labbé GM;Silk SE;Cherry CJ;Clemmensen SB;Jørgensen T;Illingworth JJ;Alanine DG;Milne KH;Ashfield R;de Jongh WA;Douglas AD;Higgins MK;Draper SJ
恶性疟原虫网织红细胞结合蛋白同源物5（PfRH 5）最近已成为针对血液期人类疟疾寄生虫的主要候选抗原。然而，已经证明鉴定可以以符合当前良好制造规范（cGMP）的方式产生基于可溶性蛋白质的疫苗的异源表达平台具有挑战性。在这里，我们报告的生产全长PfRH 5蛋白使用cGMP兼容的平台称为ExpreS 2，基于果蝇施耐德2（S2）稳定的细胞系系统。表达了PfRH 5的五种序列变体，其在去除潜在N-连接聚糖的诱变策略方面不同。所有变体结合PfRH 5受体basigin，并被一组单克隆抗体识别。免疫家兔后的分析确定了针对恶性疟原虫的功能性IgG抗体应答的定量和定性差异。由一种蛋白质变体诱导的抗体显示出与由其他疫苗平台诱导的应答定性相似。这项工作确定了果蝇S2细胞作为一个临床相关的平台，适用于生产的'难以制造'的蛋白质从疟原虫寄生虫，并确定了PfRH 5序列变异，可用于临床生产的非糖基化，可溶性全长蛋白疫苗免疫原。
The Plasmodium falciparum reticulocyte-binding protein homolog 5 (PfRH5) has recently emerged as a leading candidate antigen against the blood-stage human malaria parasite. However it has proved challenging to identify a heterologous expression platform that can produce a soluble protein-based vaccine in a manner compliant with current Good Manufacturing Practice (cGMP). Here we report the production of full-length PfRH5 protein using a cGMP-compliant platform called ExpreS2, based on a Drosophila melanogaster Schneider 2 (S2) stable cell line system. Five sequence variants of PfRH5 were expressed that differed in terms of mutagenesis strategies to remove potential N-linked glycans. All variants bound the PfRH5 receptor basigin and were recognized by a panel of monoclonal antibodies. Analysis following immunization of rabbits identified quantitative and qualitative differences in terms of the functional IgG antibody response against the P. falciparum parasite. The antibodies induced by one protein variant were shown to be qualitatively similar to responses induced by other vaccine platforms. This work identifies Drosophila S2 cells as a clinically-relevant platform suited for the production of ‘difficult-to-make’ proteins from Plasmodium parasites, and identifies a PfRH5 sequence variant that can be used for clinical production of a non-glycosylated, soluble full-length protein vaccine immunogen.