Article (Scientific journals)
Quantitation and biospecific identification of virus-like particles of human papillomavirus by capillary electrophoresis.
Bettonville, Virginie; Nicol, Jerome T. J.; Furst, Tania et al.
2017In Talanta, 175, p. 325-330
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Keywords :
Biospecific identification; Capillary electrophoresis; Papillomavirus; Quantitation
Abstract :
[en] Capillary electrophoresis (CE) for HPV-VLP quantitation is a very interesting alternative technique compared to those currently used in viral analysis, such as SDS-PAGE, Western blot or protein assay that are destructive and semi-quantitative or non specific. In this study, the quantitative performance of the CE method was evaluated. A main issue in virus quantitation is the absence of reference material. Therefore, the concentration of a HPV16-VLP sample produced in the laboratory was determined using ELISA with Gardasil(R), after adjuvant dissolution, as reference material and conformational H16.V5 antibody. HPV16-VLP concentration was found to influence particles electrophoretic mobility until a plateau was reached for concentrations </= 50microgml-1. As zeta potential is directly proportional to the electrophoretic mobility, it was measured at different HPV-VLP concentrations and the results were in complete accordance with the measured electrophoretic mobilities. The concentration dependence of the electrophoretic mobility could be explained by an overlap of the electrical double layers of adjacent particles. The HPV16-VLP peak identity was demonstrated unequivocally by the study of HPV16-VLP/H16.V5 antibody complex formation using affinity CE. Finally, the CE method was successfully validated following the ICH Q2R1 guidelines. To overcome the sample heterogeneity issue, a well-designed sample preparation was used. Considering sample complexity, validation results were satisfactory with maximum repeatability and intermediate precision RSD of 12.2% and a maximum relative bias of 1.4%.
Research center :
Centre Interfacultaire de Recherche du Médicament - CIRM
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Bettonville, Virginie ;  Université de Liège > Département de pharmacie > Analyse des médicaments
Nicol, Jerome T. J.
Furst, Tania
Thelen, Nicolas  ;  Université de Liège > Département des sciences de la vie > GIGA-R : Biologie cellulaire
Piel, Géraldine ;  Université de Liège > Département de pharmacie > Pharmacie galénique
Thiry, Marc  ;  Université de Liège > Département des sciences de la vie > GIGA-R : Biologie cellulaire
Fillet, Marianne ;  Université de Liège > Département de pharmacie > Analyse des médicaments
Jacobs, Nathalie  ;  Université de Liège > Département des sciences biomédicales et précliniques > Virologie
Servais, Anne-Catherine  ;  Université de Liège > Département de pharmacie > Analyse des médicaments
Language :
English
Title :
Quantitation and biospecific identification of virus-like particles of human papillomavirus by capillary electrophoresis.
Publication date :
2017
Journal title :
Talanta
ISSN :
0039-9140
eISSN :
1873-3573
Publisher :
Elsevier, Netherlands
Volume :
175
Pages :
325-330
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
F.R.S.-FNRS - Fonds de la Recherche Scientifique [BE]
Fonds Léon Fredericq [BE]
Commentary :
Copyright (c) 2017 Elsevier B.V. All rights reserved.
Available on ORBi :
since 28 August 2017

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