Carbon dot-based fluorescent antibody nanoprobes as brain tumour glioblastoma diagnostics.
Mattia GhirardelloRadhe ShyamXia LiuTeodoro Garcia-MillanImke SittelJavier Ramos-SorianoKathreena M KurianMaria Carmen GalanPublished in: Nanoscale advances (2022)
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.
Keyphrases
- living cells
- quantum dots
- label free
- fluorescent probe
- low cost
- resting state
- peripheral blood
- white matter
- end stage renal disease
- single molecule
- fluorescence imaging
- small molecule
- functional connectivity
- ejection fraction
- newly diagnosed
- chronic kidney disease
- cerebral ischemia
- protein protein
- randomized controlled trial
- papillary thyroid
- gene expression
- nk cells
- prognostic factors
- patient reported outcomes
- spinal cord injury
- neuropathic pain
- squamous cell carcinoma
- sensitive detection
- brain injury
- energy transfer
- subarachnoid hemorrhage
- real time pcr
- blood brain barrier
- nucleic acid
- drug discovery
- loop mediated isothermal amplification
- combination therapy
- childhood cancer