来自clade 1的未标记的肺炎球菌表面蛋白A的新方法生产和净化
Tasson da Costa Rodrigues1,2, Patricia Zorzete3, Eliane Namie Miyaji1,2
1Laboratório de Bacteriologia, Instituto Butantan, São Paulo, São Paulo, Brazil.
Applied microbiology and biotechnology
|April 3, 2024
概括
这项研究开发了一种高效的净化工艺,用于重组性肺炎球菌表面蛋白A (PspA) 来自clade 1. 这种高度纯净的PspA1可以用于新型疫苗,以提供更广泛的保护,防止Streptococcus pneumoniae感染.
科学领域:
- 微生物学 微生物学
- 疫苗学 疫苗学 疫苗学
- 蛋白质生物化学 蛋白质生物化学
背景情况:
- 肺炎链球菌会引起严重疾病,目前的疫苗由于抗体交叉活性较低,因此提供有限的血清型覆盖.
- 肺炎球菌表面蛋白A (PspA) 是一个有前途的疫苗候选人,可以提供更广泛的保护,现有研究表明PspA来自第4类 (PspA4Pro).
研究的目的:
- 开发一种高效的净化策略,用于高纯度,未标记的重组PspA,来自第1类 (PspA1).
- 为了使PspA1与PspA4Pro一起被纳入一种新型疫苗配方中,以加强肺炎球菌疾病的预防.
主要方法:
- 构建pET28a-pspA1载体和埃舍里希亚大肠杆菌BL21的转化.
- 在生物反应器中进行高密度发酵 (HDF),用于生物质生产.
- 一个多步骤的净化过程,涉及阴离子洗剂处理,阴离子交换染色学,冷沉,阴离子交换染色学和多式染色学.
主要成果:
- 实现高密度发酵,保持稳定的等离子体 (>90%) 并不形成乙酸盐.
- 开发了一种净化方案,使PSPA1具有93%的纯度,低内毒素水平,整体恢复率超过20%.
- 证明了已建立的下游加工的可扩展性和效率,用于生产未标记的PspA1.1.
结论:
- 这种新的净化工艺是有效和可扩展的,可以生产高纯度,未标记的PspA1.1.
- 这种PspA1适合纳入下一代肺炎球菌疫苗配方.
- 净化策略强调了针对PSPA变种的埋藏氨基酸的潜力.
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