Ultrasensitive Detection of Aminopeptidase N Activity in Urine and Cells with a Ratiometric Fluorescence Probe,Analytical Chemistry

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Ultrasensitive Detection of Aminopeptidase N Activity in Urine and Cells with a Ratiometric Fluorescence Probe
Analytical Chemistry ( IF 6.7 ) Pub Date : 2017-02-23 00:00:00 , DOI: 10.1021/acs.analchem.7b00021
Xinyuan He _{1,

2} , Yanhui Xu ₁ , Wen Shi _{1,

2} , Huimin Ma _{1,

2}

Affiliation

An ultrasensitive ratiometric fluorescent probe (CVN) has been designed and synthesized by incorporating alanine into the cresyl violet fluorophore. The probe shows ratiometric fluorescence response toward aminopeptidase N (APN) through the increase of fluorescent intensity ratio of 626/575 nm. The sensitivity of the probe is ultrahigh with a detection limit of 33 pg/mL, which can quantify the contents of APN in 500-fold diluted human urine samples. Furthermore, by using ratiometric fluorescence imaging, the probe reveals significantly higher contents of APN in HepG2 cells than those in LO2 cells, which has been further used to distinguish these two types of cells in mixed cocultures. The probe could be of great importance for the APN-related disease diagnosis and pathophysiology elucidation.

中文翻译：

比率荧光探针超灵敏检测尿液和细胞中氨肽酶N的活性

通过将丙氨酸掺入甲酚紫荧光团中，设计并合成了超灵敏比例荧光探针（CVN）。该探针通过增加626/575 nm的荧光强度比率显示出对氨基肽酶N（APN）的比例荧光响应。探针的灵敏度极高，检出限为33 pg / mL，可定量稀释500倍的人类尿液样品中的APN含量。此外，通过使用比例荧光成像，该探针显示出HepG2细胞中的APN含量明显高于LO2细胞中的APN含量，该探针已被进一步用于区分混合共培养中的这两种类型的细胞。该探针对于与APN相关的疾病诊断和病理生理学阐明可能非常重要。

更新日期：2017-02-23

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