识别用于净化尖端蛋白的宽谱高亲和性联体
Mengke Hu1, Xiaoyan Dong2, Qinghong Shi2
1Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, China.
Journal of chromatography. A
|April 21, 2024
概括
一种新型的,LP14,有效地净化了SARS-CoV-2的受体结合域 (RBD),包括突变变种. 这种广泛的配体显示出高亲和度和稳定的结合,对于疫苗开发和净化过程至关重要.
科学领域:
- 生物化学 生物化学
- 蛋白质净化 蛋白质净化
- 疫苗开发 疫苗开发
背景情况:
- 全球对疫苗的需求因2019年冠状病毒疾病而激增.
- 病毒突变给疫苗净化带来了挑战,特别是在亚单元疫苗中.
- 有效的净化方法对于生产安全有效的疫苗至关重要.
研究的目的:
- 开发一种用于净化SARS-CoV-2受体结合域 (RBD) 的新型亲和性.
- 选一个类库,以识别RBD的高亲缘关系联体.
- 评估已识别的对病毒突变的广泛适用性.
主要方法:
- 基于X1VX2GLNX3WX4RYSK模型生成一个612库.
- 多步骤的配体选,以识别候选.
- 分子动力学模拟来分析-RBD相互作用.
- 表面等离子体共振 (SPR) 用于确定结合亲和力.
- 使用LP14合列进行染色学净化.
主要成果:
- 确定了18种候选,其中LP14 (YVYGLNIWLRYSK) 对野生型RBD (Kd=0.520μmol/L) 具有最高的亲和力.
- 结合是由疏水性相互作用驱动的,主要的残留物已被确定.
- LP14显示病毒突变对其结合亲和力的影响很小.
- 使用LP14合色谱柱成功净化了野生型和Omicron RBD.
结论:
- LP14是一种强效的,用于RBD净化的宽谱联体.
- 鉴定的具有从复杂混合物中净化SARS-CoV-2 RBDs的显著潜力.
- 这种理性设计方法为开发用于疫苗净化的亲和联结体提供了一个有希望的策略.
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