Stefanetti Giuseppe, Saul Allan, MacLennan Calman A, Micoli Francesca
GSK Vaccines Institute for Global Health (former Novartis Vaccines Institute for Global Health NVGH) , Via Fiorentina 1, 53100 Siena, Italy.
Jenner Institute, Nuffield Department of Medicine, University of Oxford , Oxford, OX3 7DQ, United Kingdom.
Bioconjug Chem. 2015 Dec 16;26(12):2507-13. doi: 10.1021/acs.bioconjchem.5b00521. Epub 2015 Nov 18.
A solid-phase conjugation method was developed and applied to the synthesis of an O-antigen based glycoconjugate vaccine against Salmonella Typhimurium, with CRM197 as the carrier protein. Copper-free click chemistry was used as the conjugation chemistry, after derivatizing the sugar and the protein components with alkyne and azido linkers, respectively. This chemistry has the advantage of not deactivating functional groups during the conjugation step, thereby allowing the recycling of unreacted components. The activated carrier protein was adsorbed to an anion exchange matrix and quantitatively conjugated to the O-antigen. The resulting conjugate was eluted from the resin free of unconjugated sugar which was previously removed by simple washing steps. Unreacted O-antigen was recycled by addition to a new batch of resin-CRM197 resulting in further quantitative protein conjugation. This process has advantages in relation to reduction of costs for production of conjugate vaccines, allowing unreacted sugar recovery and simplifying the purification of the glycoconjugate.
开发了一种固相偶联方法，并将其应用于合成以CRM197为载体蛋白的针对鼠伤寒沙门氏菌的基于O抗原的糖缀合物疫苗。在分别用炔烃和叠氮基连接子对糖和蛋白质成分进行衍生化后，使用无铜点击化学作为偶联化学方法。这种化学方法的优点是在偶联步骤中不会使官能团失活，从而允许未反应的成分循环利用。将活化的载体蛋白吸附到阴离子交换基质上，并与O抗原进行定量偶联。所得的缀合物从树脂上洗脱下来，不含未偶联的糖，而未偶联的糖先前已通过简单的洗涤步骤除去。通过将未反应的O抗原添加到新一批的树脂-CRM197中，使其循环利用，从而实现进一步的蛋白质定量偶联。该方法在降低缀合物疫苗生产成本、实现未反应糖的回收以及简化糖缀合物的纯化方面具有优势。
