Cargando…
Carbon dot-based fluorescent antibody nanoprobes as brain tumour glioblastoma diagnostics
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 dete...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
RSC
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8962998/ https://www.ncbi.nlm.nih.gov/pubmed/35434521 http://dx.doi.org/10.1039/d2na00060a |
_version_ | 1784677896938323968 |
---|---|
author | Ghirardello, Mattia Shyam, Radhe Liu, Xia Garcia-Millan, Teodoro Sittel, Imke Ramos-Soriano, Javier Kurian, Kathreena M. Galan, M. Carmen |
author_facet | Ghirardello, Mattia Shyam, Radhe Liu, Xia Garcia-Millan, Teodoro Sittel, Imke Ramos-Soriano, Javier Kurian, Kathreena M. Galan, M. Carmen |
author_sort | Ghirardello, Mattia |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8962998 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | RSC |
record_format | MEDLINE/PubMed |
spelling | pubmed-89629982022-04-14 Carbon dot-based fluorescent antibody nanoprobes as brain tumour glioblastoma diagnostics Ghirardello, Mattia Shyam, Radhe Liu, Xia Garcia-Millan, Teodoro Sittel, Imke Ramos-Soriano, Javier Kurian, Kathreena M. Galan, M. Carmen Nanoscale Adv Chemistry 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. RSC 2022-03-03 /pmc/articles/PMC8962998/ /pubmed/35434521 http://dx.doi.org/10.1039/d2na00060a Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by/3.0/ |
spellingShingle | Chemistry Ghirardello, Mattia Shyam, Radhe Liu, Xia Garcia-Millan, Teodoro Sittel, Imke Ramos-Soriano, Javier Kurian, Kathreena M. Galan, M. Carmen Carbon dot-based fluorescent antibody nanoprobes as brain tumour glioblastoma diagnostics |
title | Carbon dot-based fluorescent antibody nanoprobes as brain tumour glioblastoma diagnostics |
title_full | Carbon dot-based fluorescent antibody nanoprobes as brain tumour glioblastoma diagnostics |
title_fullStr | Carbon dot-based fluorescent antibody nanoprobes as brain tumour glioblastoma diagnostics |
title_full_unstemmed | Carbon dot-based fluorescent antibody nanoprobes as brain tumour glioblastoma diagnostics |
title_short | Carbon dot-based fluorescent antibody nanoprobes as brain tumour glioblastoma diagnostics |
title_sort | carbon dot-based fluorescent antibody nanoprobes as brain tumour glioblastoma diagnostics |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8962998/ https://www.ncbi.nlm.nih.gov/pubmed/35434521 http://dx.doi.org/10.1039/d2na00060a |
work_keys_str_mv | AT ghirardellomattia carbondotbasedfluorescentantibodynanoprobesasbraintumourglioblastomadiagnostics AT shyamradhe carbondotbasedfluorescentantibodynanoprobesasbraintumourglioblastomadiagnostics AT liuxia carbondotbasedfluorescentantibodynanoprobesasbraintumourglioblastomadiagnostics AT garciamillanteodoro carbondotbasedfluorescentantibodynanoprobesasbraintumourglioblastomadiagnostics AT sittelimke carbondotbasedfluorescentantibodynanoprobesasbraintumourglioblastomadiagnostics AT ramossorianojavier carbondotbasedfluorescentantibodynanoprobesasbraintumourglioblastomadiagnostics AT kuriankathreenam carbondotbasedfluorescentantibodynanoprobesasbraintumourglioblastomadiagnostics AT galanmcarmen carbondotbasedfluorescentantibodynanoprobesasbraintumourglioblastomadiagnostics |