An ultrasensitive FRET-based fluorescent low molecular weight heparin nanoprobe for quantifying heparanase activity.
An ultrasensitive FRET-based fluorescent low molecular weight heparin nanoprobe for quantifying heparanase activity.
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DOI：
10.1016/j.talanta.2022.124207
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发表时间：
2023-03
期刊：
影响因子：
6.1
通讯作者：
Yayun Gu;Lizhong Peng;Weihua Ding;Yang Wang;Xuhui Zeng
中科院分区：
文献类型：
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作者：
Yayun Gu;Lizhong Peng;Weihua Ding;Yang Wang;Xuhui Zeng
肝素酶（HPA）是一种多肽-β-葡萄糖醛酸酶，它的失调会促进癌症的转移。开发快速、灵敏地监测HPA酶活性的技术对于寻找针对HPA的分子治疗方法至关重要。在此，我们开发了一种基于荧光共振能量转移（FRET）的新型纳米探针AuNCs-LMWH-AuNRs，将AuNCs@GSH-cys和AuNRs/端- nh2 /侧- sio2分别连接到低分子量肝素（LMWH）的非还原端和还原端。AuNCs@GSH-cys的绝对量子产率为1.1%。AuNRs/end-NH2/side-SiO2的吸收光谱（最大纵向吸收825 nm）与AuNCs@GSH-cys的发射光谱（最大发射824 nm）精确重合，进一步提高了FRET的效率。在HPA存在下，LMWH纳米探针表现出超灵敏的响应，激发/发射波长(lambda （ex) = 560 nm, lambda (em) = 824 nm）。该探针的线性动态检测范围为0.125 ng/μL ~ 0.01 μg/μLin，检出限为82.15 pM （0.43 pg/μL）。该探针具有良好的选择性和荧光开启效率，可一步检测细胞肝素酶活性。高效的低分子肝素纳米探针也可以实现HPA抑制剂的高通量筛选。
Heparanase (HPA) is a multifacetedendo-β-glucuronidase, and its dysregulation facilitates cancer metastasis. Developing techniques for fast and sensitively monitoring HPA enzymatic activity is crucial for searching for molecular therapies targeting HPA. Herein, we developed a novel fluorescence resonance energy transfer (FRET)-based nanoprobe AuNCs-LMWH-AuNRs, with AuNCs@GSH-cys and AuNRs/end-NH2/side-SiO2attached to the non-reducing terminus and reducing terminus of low molecular weight heparin (LMWH), respectively. AuNCs@GSH-cys exhibited an absolute quantum yield of 1.1%. The absorption spectra of AuNRs/end-NH2/side-SiO2(825 nm for maximum longitudinal absorption) and the emission spectra of AuNCs@GSH-cys (824 nm for maximum emission) were precisely overlapping, further enhancing the efficiency of FRET. In the presence of HPA, the LMWH nanoprobe exhibited an ultrasensitive response with excitation/emission wavelength (lambda (ex) = 560 nm, lambda (em) = 824 nm). The probe presented a wide linear dynamic detection range (LDR) of 0.125 ng/μL - 0.01 μg/μLin vitrowith a limit of detection (LODs) of 82.15 pM (0.43 pg/μL). The excellent selectivity and good fluorescence turn-on efficiency of the probe made it possible for one-step detection of cellular heparanase activity. High throughput screening of HPA inhibitors also can be accomplished using the highly efficient LMWH nanoprobe.