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Comparative Evaluation of Two Methods for Preparative Fractionation of Proteinaceous Subvisible Particles-Differential Centrifugation and FACS

Boll, Björn and Folzer, Emilien and Finkler, Christof and Huwyler, Jörg and Mahler, Hanns-Christian and Schmidt, Roland and Koulov, Atanas V.. (2015) Comparative Evaluation of Two Methods for Preparative Fractionation of Proteinaceous Subvisible Particles-Differential Centrifugation and FACS. Pharmaceutical research, 32 (12). pp. 3952-3964.

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Official URL: http://edoc.unibas.ch/40936/

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Abstract

The goal of this study was to compare and evaluate two preparative techniques for fractionation of proteinaceous subvisible particles. This work enables future studies to address the potential biological consequences of proteinaceous subvisible particles in protein therapeutic products.
Particles were generated by heat stress and separated by size using differential centrifugation and FACS (Fluorescence-activated cell sorter). Resulting fractions were characterized by size-exclusion chromatography, light obscuration, flow imaging microscopy and resonant mass measurement.
Here we report the optimization and comprehensive evaluation of two methods for preparative fractionation of subvisible proteinaceous particles into distinct size fractions in the range between 0.25 and 100 mu m: differential centrifugation and FACS. Using these methods, well-defined size fractions were prepared and characterized in detail. Critical assessment and comparison of the two techniques demonstrated their complementarity and for the first time-their relative advantages and drawbacks.
FACS and differential centrifugation are valuable tools to prepare well-defined size-fractions of subvisible proteinaceous particles. Both techniques possess unique and advantageous attributes and will likely find complementary application in future research on the biological consequences of proteinaceous subvisible particles.
Faculties and Departments:05 Faculty of Science > Departement Pharmazeutische Wissenschaften > Pharmazie > Pharmaceutical Technology (Huwyler)
UniBasel Contributors:Huwyler, Jörg and Boll, Björn and Folzer, Emilien
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Springer
ISSN:0724-8741
e-ISSN:1573-904X
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:11 Aug 2016 08:18
Deposited On:11 Aug 2016 08:18

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