Hybrid Macromolecular Constructs as a Platform for Spectral Nanoprobes for Advanced Cellular Barcoding in Flow Cytometry.
antibody-polymer-dye conjugates
cellular barcoding
double fluorescence
flow cytometry
nanoprobes
Journal
Macromolecular bioscience
ISSN: 1616-5195
Titre abrégé: Macromol Biosci
Pays: Germany
ID NLM: 101135941
Informations de publication
Date de publication:
11 Sep 2023
11 Sep 2023
Historique:
revised:
17
08
2023
received:
29
06
2023
pubmed:
11
9
2023
medline:
11
9
2023
entrez:
11
9
2023
Statut:
aheadofprint
Résumé
Herein, an advanced bioconjugation technique to synthesize hybrid polymer-antibody nanoprobes tailored for fluorescent cell barcoding in flow cytometry-based immunophenotyping of leukocytes is applied. A novel approach of attachment combining two fluorescent dyes on the copolymer precursor and its conjugation to antibody is employed to synthesize barcoded nanoprobes of antibody polymer dyes allowing up to six nanoprobes to be resolved in two-dimensional cytometry analysis. The major advantage of these nanoprobes is the construct design in which the selected antibody is labeled with an advanced copolymer bearing two types of fluorophores in different molar ratios. The cells after antibody recognition and binding to the target antigen have a characteristic double fluorescence signal for each nanoprobe providing a unique position on the dot plot, thus allowing antibody-based barcoding of cellular samples in flow cytometry assays. This technique is valuable for cellular assays that require low intersample variability and is demonstrated by the live cell barcoding of clinical samples with B cell abnormalities. In total, the samples from six various donors were successfully barcoded using only two detection channels. This barcoding of clinical samples enables sample preparation and measurement in a single tube.
Identifiants
pubmed: 37691533
doi: 10.1002/mabi.202300306
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e2300306Subventions
Organisme : Ministry of Health of the Czech Republic
ID : NU21-08-00280
Organisme : Ministry of Health of the Czech Republic
ID : NU23J-03-00026
Organisme : Ministry of Industry and Trade of the Czech Republic
ID : FV10370
Organisme : European Union
Organisme : Ministerstvo Školství, Mládeže a Tělovýchovy
ID : LX22NPO5102
Informations de copyright
© 2023 The Authors. Macromolecular Bioscience published by Wiley-VCH GmbH.
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