Carbon Dot-based Fluorescent Antibody Nanoprobes as Brain Tumour Glioblastoma Diagnostics
Carbon Dot-based Fluorescent Antibody Nanoprobes as Brain Tumour Glioblastoma Diagnostics
复制标题
基于碳点的荧光抗体纳米探针用于脑肿瘤胶质母细胞瘤诊断
DOI：
10.1101/2021.11.29.470408
复制
发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Ghirardello M
中科院分区：
文献类型：
--
作者：
Ghirardello M
开发用于检测患者脑癌的有效和灵敏的工具至关重要，特别是因为许多这些肿瘤直到疾病进展和治疗效果不佳时才被诊断出来。目前的策略采用抗体（Abs）来检测组织样品中的胶质纤维酸性蛋白（GFAP），因为GFAP是大脑所特有的，并且不存在于正常外周血中，并且它依赖于荧光报告子。在此，我们描述了一种低成本、实用和通用的方法，用于用荧光碳量子点（CD）标记蛋白质和抗体，以产生稳健、光稳定且适用于临床环境的诊断探针。两步方案依赖于通过将酰胺缀合和应变促进的炔-叠氮化物环加成（SPAAC）连接化学结合来使二苯并环辛炔（DBCO）官能化CD与叠氮化物官能化蛋白质缀合。使用这种策略开发的新类别的Ab-CD缀合物被成功地用于验证该方法的胶质母细胞瘤（GBM）患者的人脑组织的免疫组织化学染色。总的来说，这些新型荧光探针在成本、光稳定性和适用性方面提供了一种有前途的通用策略，可以扩展到其他抗体和蛋白质系统。
The development of efficient and sensitive tools for the detection of brain cancer in patients is of the utmost importance particularly because many of these tumours go undiagnosed until the disease has advanced and when treatment is less effective. Current strategies employ antibodies (Abs) to detect Glial Fibrillary Acid Protein (GFAP) in tissue samples, since GFAP is unique to the brain and not present in normal peripheral blood, and it relies on fluorescent reporters. Herein we describe a low cost, practical and general method for the labelling of proteins and antibodies with fluorescent carbon dots (CD) to generate diagnostic probes that are robust, photostable and applicable to the clinical setting. The two-step protocol relies on the conjugation of a dibenzocyclooctyne (DBCO)-functionalised CD with azide functionalised proteins by combining amide conjugation and strain promoted alkyne–azide cycloaddition (SPAAC) ligation chemistry. The new class of Ab-CD conjugates developed using this strategy was successfully used for the immunohistochemical staining of human brain tissues of patients with glioblastoma (GBM) validating the approach. Overall, these novel fluorescent probes offer a promising and versatile strategy in terms of costs, photostability and applicability which can be extended to other Abs and protein systems.