用实验设计控制来缓解商业传感器芯片变化的策略.
Eliza K Hanson1,2, Chien-Wei Wang1,2, Lisa Minkoff2
1Department of Chemistry, University of Kansas, Lawrence, KS 66047, USA.
Sensors (Basel, Switzerland)
|August 12, 2023
概括
这项研究确定了使用Ni-NTA芯片用于生物分子相互作用的表面等离子体共振 (SPR) 信号变异的因素. 建议采用校准和规范化策略,以提高SPR实验中的数据可重现性和质量.
科学领域:
- 生物化学 生物化学
- 分析化学 分析化学
- 生物技术是生物技术.
背景情况:
- 表面等离子共振 (SPR) 是一种关键的实时生物传感技术,用于测量结合动力学和亲和力.
- 尼-NTA固定是SPR中联体附着的常见方法,提供传感器再生,但可能导致信号变化.
- 使用Ni-NTA芯片的SPR实验中的可重复性挑战阻碍了可靠的数据生成.
研究的目的:
- 调查导致Ni-NTA SPR传感器芯片信号变化的因素.
- 制定提高SPR实验可重现性和数据质量的策略.
- 解决关于商用SPR传感器芯片的变异性的知识差距.
主要方法:
- 利用尼科亚开放SPR系统作为研究SPR变异性的模型.
- 采用卵巢癌生物标记蛋白 (MUC16,HE4) 和它们的抗体作为模型配体/分析物对.
- 评估了非特异性结合,芯片对芯片的可变性,最大固定化差异,以及对分析物响应的配体密度影响.
主要成果:
- 在多个Ni-NTA芯片中观察到显著的非特异性结合.
- 与不同芯片相比,在同一芯片上显示出更高的连接体固定和分析物结合的一致性.
- 确定了不同的最大固定能力和连接体密度和分析物反应之间的关系,归因于固态拥挤.
结论:
- 芯片对芯片的可变性和连接体固定密度是影响SPR数据可重现性的关键因素.
- 基于连接体不动化水平的校准和规范化可以显著改善数据质量并最大限度地降低变化.
- 在实验设计中实施这些策略可以提高SPR传感平台对生物分子相互作用分析的可靠性.
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