English   español  
Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/219704
Share/Impact:
Statistics
logo share SHARE logo core CORE   Add this article to your Mendeley library MendeleyBASE

Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL | DATACITE
Exportar a otros formatos:

Title

Lot-to-lot stability of antibody reagents for flow cytometry

AuthorsBöttcher, Sebastian; Velden, Vincent H. J. van der; Villamor, Neus; Ritgen, Matthias; Flores-Montero, Juan; Murua Escobar, Hugo; Kalina, Tomas; Brüggemann, Monika; Grigore, Georgiana Emilia; Martín-Ayuso, Marta; Lécrevisse, Quentin; Pedreira, C. E.; Dongen, J. J. M. van; Orfao, Alberto
KeywordsFluorescence intensity
Variability
Fluorochrome-to-antibody ratio
Flow cytometry
Issue Date2019
PublisherElsevier
CitationJournal of Immunological Methods 475: 112294 (2019)
AbstractThe fluorescence detected using fluorochrome-labelled monoclonal antibodies depends not only on the abundance of the target antigen, but amongst many other factors also on the effective fluorochrome-to-antibody ratio. The diagnostic approach of the EuroFlow consortium relies on reproducible fluorescence intensities over time. A capture bead system for mouse immunoglobulin light chains was utilized to compare the mean fluorescence intensity of 1323 consecutive antibody lots to the currently used lot of the same monoclonal antibody. In total, 157 different monoclonal antibodies were assessed over seven years. Median relative difference between consecutive lots was 3.8% (range: 0.01% to 164.7%, interquartile range: 1.3% to 10.1%). The relative difference exceeded 20% in 8.8% of all comparisons. FITC labelled monoclonal antibodies (median relative difference: 2.1%) showed a significantly smaller variation between lots than antibodies conjugated to PE (3.5%), PECy7 (3.9%), PerCPCy5.5 (5.8%), APC (5.8%), APCH7 (7.4%), and APCC750 (14.5%). Reagents labelled with Pacific Blue (1.4%), Pacific Orange (2.4%), HV450 (0.7%), and HV500 (1.7%) demonstrated more consistent results compared to conjugates of BV421 (4.1%) and BV510 (16.2%). Additionally, significant differences in lot-to-lot fluorescence stability amongst antibodies labelled with the same fluorochrome were observed between manufacturers. These observations might guide future quality recommendations for the production and application of fluorescence-labelled monoclonal antibodies in multicolor flow cytometry.
Publisher version (URL)https://doi.org/10.1016/j.jim.2017.03.018
URIhttp://hdl.handle.net/10261/219704
DOIhttp://dx.doi.org/10.1016/j.jim.2017.03.018
ISSN0022-1759
Appears in Collections:(IBMCC) Artículos
Files in This Item:
File Description SizeFormat 
accesoRestringido.pdf59,24 kBAdobe PDFThumbnail
View/Open
Show full item record
Review this work
 


WARNING: Items in Digital.CSIC are protected by copyright, with all rights reserved, unless otherwise indicated.