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一个双酶放大循环,用于敏感的生物传感内酶

Chuanwen Zhou¹, Xiaomin Li¹, Sze Wing Tang¹

¹Department of Chemistry, City University of Hong Kong, Tat Chee Avenue, Kowloon, Hong Kong.

|July 24, 2023

概括

一个新的生物传感器使用分析剂触发的链接器固定酶 (AMELIE) 相互解放机制来快速和灵敏地检测内酶. 这种方法实现了对原酶的低检测极限,可以在培养物中识别细菌原酶.

科学领域:

生物化学和生物技术
酶检测仪和生物传感器
分析化学分析化学

背景情况:

内酶在生物过程和疾病中起着至关重要的作用.
对内酶的敏感和快速检测方法对于研究和诊断至关重要.
现有的方法可能缺乏敏感性,速度或适应性,以适应各种内酶.

研究的目的:

开发一种新的生物传感器平台,用于敏感的内酶检测.
为了证明分析剂触发的链接器固定酶 (AMELIE) 机制的相互解放.
为了验证AMELIE生物传感器使用原酶作为模型分析剂.

主要方法:

开发了AMELIE机制,其中涉及一对自我切割,链接器固定酶.
采用了原酶-酸酶对和颜色对比的卜过氧化酶反应来进行信号传导.
通过特定的链接器 (酸盐和链) 固定化酶,由目标分析物或其他酶分裂.

主要成果:

达到了2.5 pg mL-1的高灵敏度检测极限,用于4-log线性范围的原酶.
在细菌培养超级生物中成功检测出细胞外原酶.
对大肠杆菌 (E. coli) 的敏感度低至103个殖民地形成单位mL-1.

结论:

艾米利机制为内酶检测提供了一个快速,灵敏和可适应的平台.
开发的生物传感器显示出在复杂的生物样本中检测酶的巨大潜力.
艾米利技术可以很容易地被修改为检测各种其他内酶.

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