Development of an O-polysaccharide based recombinant glycoconjugate vaccine in engineered E. coli against ExPEC O1
Development of an O-polysaccharide based recombinant glycoconjugate vaccine in engineered E. coli against ExPEC O1
复制标题
在工程大肠杆菌中开发针对 ExPEC O1 的基于 O-多糖的重组糖复合物疫苗
DOI：
10.1016/j.carbpol.2021.118796
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发表时间：
2021-11-09
影响因子：
11.2
通讯作者：
Feng, Lu
中科院分区：
文献类型：
--
作者：
Jiang, Xiaolong;Bai, Jing;Feng, Lu
肠外致病性大肠杆菌O1是一种常见的血清型，可引起严重感染，并且通常对抗菌治疗无效。糖缀合物疫苗代表了减少ExPEC感染的有希望的措施。在此，我们设计了O1特异性糖优化的底盘菌株用于生产O多糖（OPS）抗原和基于OPS的生物缀合物。具体地，当与亲本菌株的不可测量的水平相比时，在糖优化的底盘菌株中合成OPS和基于OPS的糖蛋白。寡糖基转移酶和载体蛋白的优化表达进一步提高了效价。质谱分析表明，在载体蛋白的常规和未报道的糖基化序列处，得到的生物缀合物结构正确，具有较高的糖基化效率。最后，纯化的生物缀合物刺激小鼠产生特异性IgG抗体，并保护它们免受强毒ExPEC O1攻击。该即插即用的糖优化平台适用于生物缀合物合成，从而为未来的医学应用提供了潜在的平台。
Extraintestinal pathogenic Escherichia coli O1 is a frequently identified serotype that causes serious infections and is often refractory to antimicrobial therapy. Glycoconjugate vaccine represents a promising measure to reduce ExPEC infections. Herein, we designed an O1-specific glyco-optimized chassis strain for manufacture of Opolysaccharide (OPS) antigen and OPS-based bioconjugate. Specifically, OPS and OPS-based glycoprotein were synthesized in glyco-optimized chassis strain, when compared to the unmeasurable level of the parent strain. The optimal expression of oligosaccharyltransferase and carrier protein further improved the titer. MS analysis elucidated the correct structure of resulting bioconjugate at routine and unreported glycosylation sequons of carrier protein, with a higher glycosylation efficiency. Finally, purified bioconjugate stimulated mouse to generate specific IgG antibodies and protected them against virulent ExPEC O1 challenge. The plug-and-play glyco-optimized platform is suitable for bioconjugate synthesis, thus providing a potential platform for future medical applications.