Costa M H, Sant'Anna O A, de Araujo P S, Sato R A, Quintilio W, Silva L V, Matos C R, Raw I
Laboratório de Microesferas e lipossomos-Centro de Biotecnologia, Butantan, Brasil.
Appl Biochem Biotechnol. 1998 Apr;73(1):19-28. doi: 10.1007/BF02788830.
Protein stability is one of the most important obstacles for successful formulation in the development of new-generation vaccines. Here, the 18kDa heat-shock protein (18kDa-hsp) was chemically modified though conjugation with bovine serum albumin or by esterification with N-hydroxysuccinimide ester of palmitic acid. The biologically active conformation of the protein was preserved after chemical modification. The immune responses to the recombinant 18kDa-hsp from Mycobacterium leprae were studied in different presentations: free, copolymerized with bovine serum albumin in aggregates (18kDa-hsp-BSA), and either surface linked to liposomes or entrapped into liposomes. Measuring the antibody production of immunized genetically selected mice has compared the adjuvant effects of liposomes and proteic copolymer. Among the two liposome preparations, the strongest response was obtained with the surface-exposed antigen-liposomes. The copolymer 18kDa-hsp-BSA conferred a high titer of antibody in injected mice, and persisted 70 d after immunization. This approach should prove very useful for designing more effective vaccines by using 18kDa-hsp as carrier protein.
蛋白质稳定性是新一代疫苗研发中成功制剂的最重要障碍之一。在此，通过与牛血清白蛋白偶联或用棕榈酸的N-羟基琥珀酰亚胺酯酯化对18kDa热休克蛋白（18kDa-hsp）进行化学修饰。化学修饰后蛋白质的生物活性构象得以保留。对来自麻风分枝杆菌的重组18kDa-hsp的免疫反应在不同形式下进行了研究：游离形式、与牛血清白蛋白共聚形成聚集体（18kDa-hsp-BSA）、表面连接到脂质体或包封在脂质体中。通过测量免疫的基因选择小鼠的抗体产生，比较了脂质体和蛋白质共聚物的佐剂效果。在两种脂质体制剂中，表面暴露抗原的脂质体产生的反应最强。共聚物18kDa-hsp-BSA在注射小鼠中产生了高滴度抗体，并在免疫后持续70天。这种方法对于利用18kDa-hsp作为载体蛋白设计更有效的疫苗应该非常有用。
