A Robust Method for Assaying the Immunoreactive Fraction in Nonequilibrium Systems.

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Etat: Public
Version: de l'auteur⸱e
Licence: CC BY 4.0
ID Serval
serval:BIB_E07AD7079702
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
A Robust Method for Assaying the Immunoreactive Fraction in Nonequilibrium Systems.
Périodique
Pharmaceuticals
Auteur⸱e⸱s
Denoël T., Pedrelli L., Pantaleo G., Prior J.O.
ISSN
1424-8247 (Print)
ISSN-L
1424-8247
Statut éditorial
Publié
Date de publication
03/12/2019
Peer-reviewed
Oui
Volume
12
Numéro
4
Pages
E177
Langue
anglais
Notes
Publication types: Journal Article
Publication Status: epublish
Résumé
The immunoreactive fraction r provides important information on the functional purity of radiolabeled proteins. It is traditionally determined by saturating the radioimmunoconjugate with an increasing excess of antigen, followed by linear extrapolation to infinite antigen excess in a double inverse "Lindmo plot". Although several reports have described shortcomings in the Lindmo plot, a systematic examination is lacking. Using an experimental and simulation-based approach, we compared-for accuracy, precision and robustness-the Lindmo plot with the "rectangular hyperbola" extrapolation method based on the Langmuir model. The differences between the theoretical and extrapolated r values demonstrate that nonequilibrium and antigen depletion are important sources of error. The mathematical distortions resulting from the linearization of the data in the Lindmo plot induce fragility towards stochastic errors and make it necessary to exclude low bound fractions. The rectangular hyperbola provides robust and precise r estimates from raw binding data, even for slow kinetics.
Mots-clé
Lindmo plot, binding sites, antibody, immunoconjugates, immunoreactivity, radiolabeled antibodies, radiopharmaceuticals, translational pharmacology
Pubmed
Web of science
Open Access
Oui
Création de la notice
15/12/2019 17:18
Dernière modification de la notice
28/01/2020 6:20
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