Disclosed are methods, systems, components, and compositions for cell-free synthesis of glycosylated carrier proteins. The glycosylated carrier proteins may be utilized in vaccines, including anti-bacterial vaccines. The glycosylated carrier proteins may include a bacterial polysaccharide conjugated to a carrier, which may be utilized to generate an immune response in an immunized host against the polysaccharide conjugated to the carrier. The glycosylated carrier proteins may be synthesized in cell-free glycoprotein synthesis (CFGpS) systems using prokaryote cell lysates that are enriched in components for glycoprotein synthesis such as oligosaccharyltransferases (OSTs) and lipid-linked oligosaccharides (LLOs) including OSTs and LLOs associated with synthesis of bacterial O antigens.
所公开的是无细胞合成糖基化载体蛋白的方法、系统、组件和组合物。糖基化载体蛋白可用于疫苗,包括抗菌疫苗。糖基化载体蛋白可包括与载体共轭的细菌
多糖,可用于在免疫宿主中产生针对与载体共轭的
多糖的免疫反应。糖基化载体蛋白可在无细胞糖蛋白合成(CFGpS)系统中合成,该系统使用富含糖蛋白合成成分的原核细胞裂解物,如低聚糖转移酶(OST)和脂联低聚糖(LLO),包括与合成细菌 O 抗原有关的 OST 和 LLO。