SERS nanotags for folate receptor a detection at the single cell level: discrimination of overexpressing cells and potential for live cell applications.
SERS nanotags for folate receptor a detection at the single cell level: discrimination of overexpressing cells and potential for live cell applications.
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用于叶酸受体的 SERS 纳米标签在单细胞水平上进行检测：过度表达细胞的区分和活细胞应用的潜力。
DOI：
10.1039/d2an00706a
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Verdin A
中科院分区：
文献类型：
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作者：
Verdin A
叶酸受体α（FRα）是一种高亲和力的叶酸膜受体，在多种癌症中过度表达。检测该受体的过表达对于癌细胞鉴定和潜在地指导治疗的选择是重要的，因为几种FRα靶向药物目前正在临床试验中。在这项工作中，我们建立了Sers纳米标签的基础上核@壳Au@Ag纳米粒子标记共振拉曼报告和功能化的硫醇化PEG连接器轴承叶酸在链端。使用单细胞上的Sers图谱，我们表明纳米标签（FR-纳米标签）可以特异性地靶向过表达HeLa细胞上的FRα，并且可以测量沿着纳米标签沿着引入培养基中的游离叶酸对FRα的逐渐阻断。使用对照纳米标签，我们表明，与没有叶酸的PEG链相比，当叶酸存在于PEG接头上时，HeLa细胞上的Sers响应高10倍。FR-纳米标签的非特异性结合被证明是低的，并且主要由PEG链末端的叶酸分子引起。当比较具有不同FRα表达水平的癌细胞时，我们在过表达的HeLa细胞上获得了比非过表达的A549细胞高4倍的Sers响应，从而能够以高对比度区分两种细胞系。由于所开发的纳米标签的生物相容性，我们证明了在活的HeLa细胞上测量FRα也是可能的，并且给出了与固定细胞上的测量类似的结果，表明所开发的纳米标签在各种实验条件下用于检测FRα的通用性。
Folate receptor α (FRα) is a high affinity folate membrane receptor that is overexpressed in a wide variety of cancers. Detecting the overexpression of this receptor is important for cancer cells identification and to potentially guide the choice of treatment since several FRα-targeted drugs are currently in clinical trials. In this work, we built SERS nanotags based on core@shell Au@Ag nanoparticles labelled with resonant Raman-reporter and functionalised with a thiolated PEG linker bearing folic acid at the chain end. Using SERS mapping on single cells, we showed that the nanotags (FR-nanotags) could specifically target FRα on overexpressing HeLa cells and could measure the gradual blocking of FRα by free folic acid introduced in the media along the nanotags. With a control nanotag, we showed that the SERS response was 10-fold higher on HeLa cells when folic acid is present on the PEG linker compared to PEG chains without folic acid. Non-specific binding of the FR-nanotags was demonstrated to be low and mainly caused by the folic acid molecule at the PEG chain end. When comparing cancer cells with different expression levels of FRα, we obtained 4-fold higher SERS response on overexpressing HeLa cells compared to non-overexpressing A549 cells, allowing the discrimination of both cell lines with a high contrast. Owing to the biocompatibility of the developed nanotags, we demonstrated that measurements of FRα on live HeLa cells were also possible and gave similar results to measurements on fixed cells, indicating the versatility of the developed nanotags for detecting FRα under various experimental conditions.