Combined Chemical Synthesis and Tailored Enzymatic Elongation Provide Fully Synthetic and Conjugation-Ready Neisseria meningitidis Serogroup X Vaccine Antigens,ACS Chemical Biology

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Combined Chemical Synthesis and Tailored Enzymatic Elongation Provide Fully Synthetic and Conjugation-Ready Neisseria meningitidis Serogroup X Vaccine Antigens
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2018-02-26 00:00:00 , DOI: 10.1021/acschembio.7b01057
Davide Oldrini ₁ , Timm Fiebig ₂ , Maria Rosaria Romano ₁ , Daniela Proietti ₁ , Monika Berger ₂ , Marta Tontini ₁ , Riccardo De Ricco ₁ , Laura Santini ₁ , Laura Morelli ₃ , Luigi Lay ₃ , Rita Gerardy-Schahn ₂ , Francesco Berti ₁ , Roberto Adamo ₁

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Studies on the polymerization mode of Neisseria meningitidis serogroup X capsular polymerase CsxA recently identified a truncated construct that can be immobilized and used for length controlled on-column production of oligosaccharides. Here, we combined the use of a synthetic acceptor bearing an appendix for carrier protein conjugation and the on-column process to a novel chemo-enzymatic strategy. After protein coupling of the size optimized oligosaccharide produced by the one-pot elongation procedure, we obtained a more homogeneous glycoconjugate compared to the one previously described starting from the natural polysaccharide. Mice immunized with the conjugated fully synthetic oligomer elicited functional antibodies comparable to controls immunized with the current benchmark MenX glycoconjugates prepared from the natural capsule polymer or from fragments of it enzymatically elongated. This pathogen-free technology allows the fast total in vitro construction of predefined bacterial polysaccharide fragments. Compared to conventional synthetic protocols, the procedure is more expeditious and drastically reduces the number of purification steps to achieve the oligomers. Furthermore, the presence of a linker for conjugation in the synthetic acceptor minimizes manipulations on the enzymatically produced glycan prior to protein conjugation. This approach enriches the methods for fast construction of complex bacterial carbohydrates.

中文翻译：

化学合成和量身定制的酶促延伸相结合，可提供完全合成和结合的脑膜炎奈瑟氏球菌血清群X疫苗抗原。

脑膜炎奈瑟菌的聚合方式研究血清群X荚膜聚合酶CsxA最近鉴定出一种可固定的截短构建体，可用于长度控制的柱上寡糖生产。在这里，我们结合使用带有附件的合成受体来进行载体蛋白的缀合和柱上过程成为一种新型的化学酶促策略。在通过一锅延伸程序产生的大小优化的寡糖进行蛋白质偶联后，与从天然多糖开始的前述糖偶联物相比，我们获得了更均一的糖偶联物。用缀合的完全合成的寡聚物免疫的小鼠引发的功能抗体可与用目前的基准MenX糖缀合物免疫的对照相比，后者是由天然胶囊聚合物或其酶促酶切的片段制备的。体外构建预定的细菌多糖片段。与常规合成方案相比，该过程更加快捷，并大大减少了获得低聚物的纯化步骤。此外，在合成受体中用于缀合的接头的存在最小化了在蛋白质缀合之前对酶促产生的聚糖的操纵。这种方法丰富了快速构建复杂细菌碳水化合物的方法。

更新日期：2018-02-26

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