caa a22 4500 445312335 CHVBK 20180317142634.0 cr unu---uuuuu 170323e20110701xx s 000 0 eng 10.1007/s11095-011-0393-x doi (NATIONALLICENCE)springer-10.1007/s11095-011-0393-x High-Throughput Screening of Excipients Intended to Prevent Antigen Aggregation at Air-Liquid Interface [Elektronische Daten] [Sébastien Dasnoy, Nancy Dezutter, Dominique Lemoine, Vivien Le Bras, Véronique Préat] ABSTRACT: Purpose: The aim was to develop a high-throughput screening method compatible with low protein concentrations, as present in vaccines, in order to evaluate the performance of various excipients in preventing the aggregation at air-liquid interface of an experimental recombinant antigen called Antigen 18A. Methods: Aggregation of Antigen 18A was triggered by shaking in a half-filled vial or by air bubbling in a microplate. Size-exclusion chromatography, turbidimetry, Nile Red fluorescence spectroscopy, and attenuated total reflection Fourier-transform infrared spectroscopy were used to assess Antigen 18A aggregation. A high-throughput method, based on tryptophan fluorescence spectroscopy, was set up to screen excipients for their capability to prevent Antigen 18A aggregation at air-liquid interface. Results: While a similar aggregation profile was obtained with both stress tests when using size-exclusion chromatography, spectroscopic and turbidimetric methods showed an influence of the stress protocol on the nature of the aggregates. The high-throughput screening revealed that 7 out of 44 excipients significantly prevented Antigen18A from aggregating. We confirmed the performance of hydroxypropyl-β-cyclodextrin and hydroxypropyl-γ-cyclodextrin, as well as poloxamers 188 and 407, in half-filled shaken vials. Conclusions: A high-throughput screening approach can be followed for evaluating the performance of excipients against aggregation of a protein antigen at air-liquid interface. Springer Science+Business Media, LLC, 2011 excipients nationallicence fluorescence spectroscopy nationallicence high-throughput screening nationallicence protein aggregation nationallicence vaccines nationallicence Dasnoy Sébastien Louvain Drug Research Institute, Université catholique de Louvain, Unité de Pharmacie Galénique, Avenue E. Mounier 73, UCL 7320, 1200, Brussels, Belgium aut Dezutter Nancy GlaxoSmithKline Biologicals, Rue de l'Institut 89, 1330, Rixensart, Belgium aut Lemoine Dominique GlaxoSmithKline Biologicals, Rue de l'Institut 89, 1330, Rixensart, Belgium aut Le Bras Vivien GlaxoSmithKline Biologicals, Rue de l'Institut 89, 1330, Rixensart, Belgium aut Préat Véronique Louvain Drug Research Institute, Université catholique de Louvain, Unité de Pharmacie Galénique, Avenue E. Mounier 73, UCL 7320, 1200, Brussels, Belgium aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/7(2011-07-01), 1591-1605 0724-8741 28:7<1591 2011 28 11095 https://doi.org/10.1007/s11095-011-0393-x text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-011-0393-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Dasnoy Sébastien Louvain Drug Research Institute, Université catholique de Louvain, Unité de Pharmacie Galénique, Avenue E. Mounier 73, UCL 7320, 1200, Brussels, Belgium aut NATIONALLICENCE 700 1- Dezutter Nancy GlaxoSmithKline Biologicals, Rue de l'Institut 89, 1330, Rixensart, Belgium aut NATIONALLICENCE 700 1- Lemoine Dominique GlaxoSmithKline Biologicals, Rue de l'Institut 89, 1330, Rixensart, Belgium aut NATIONALLICENCE 700 1- Le Bras Vivien GlaxoSmithKline Biologicals, Rue de l'Institut 89, 1330, Rixensart, Belgium aut NATIONALLICENCE 700 1- Préat Véronique Louvain Drug Research Institute, Université catholique de Louvain, Unité de Pharmacie Galénique, Avenue E. Mounier 73, UCL 7320, 1200, Brussels, Belgium aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/7(2011-07-01), 1591-1605 0724-8741 28:7<1591 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer