Department of Chemistry, University of Florence, Via della Lastruccia 3, 50019, Sesto Fiorentino, Italy.
Magnetic Resonance Center (CERM), University of Florence and Consorzio Interuniversitario Risonanze Magnetiche di Metallo Proteine (CIRMMP), Via L. Sacconi 6, 50019, Sesto Fiorentino, Italy.
Angew Chem Int Ed Engl. 2017 Nov 20;56(47):14997-15001. doi: 10.1002/anie.201709274. Epub 2017 Oct 19.
Carbohydrate-based vaccines are among the safest and most effective vaccines and represent potent tools for prevention of life-threatening bacterial infectious diseases, like meningitis and pneumonia. The chemical conjugation of a weak antigen to protein as a source of T-cell epitopes generates a glycoconjugate vaccine that results more immunogenic. Several methods have been used so far to characterize the resulting polysaccharide-protein conjugates. However, a reduced number of methodologies has been proposed for measuring the degree of saccharide conjugation at the possible protein sites. Here we show that detailed information on large proteins conjugated with large polysaccharides can be achieved by a combination of solution and solid-state NMR spectroscopy. As a test case, a large protein assembly, l-asparaginase II, has been conjugated with Neisseria meningitidis serogroup C capsular polysaccharide and the pattern and degree of conjugation were determined.
基于碳水化合物的疫苗是最安全和最有效的疫苗之一，是预防危及生命的细菌性传染病（如脑膜炎和肺炎）的有力工具。将弱抗原与蛋白质化学缀合作为 T 细胞表位的来源，可产生更具免疫原性的糖缀合物疫苗。迄今为止，已经使用了几种方法来表征所得多糖-蛋白质缀合物。然而，目前提出的用于测量可能的蛋白质部位上糖缀合程度的方法数量有限。在这里，我们展示了通过溶液和固态 NMR 光谱学的组合，可以获得与大多糖缀合的大蛋白质的详细信息。作为一个测试案例，已将大型蛋白质组装体 II 型天冬酰胺酶与脑膜炎奈瑟菌 C 群荚膜多糖缀合，并确定了缀合的模式和程度。
