caa a22 4500 445313862 CHVBK 20180317142639.0 cr unu---uuuuu 170323e20110501xx s 000 0 eng 10.1007/s11095-011-0374-0 doi (NATIONALLICENCE)springer-10.1007/s11095-011-0374-0 Fluorescence Single Particle Tracking for the Characterization of Submicron Protein Aggregates in Biological Fluids and Complex Formulations [Elektronische Daten] [Vasco Filipe, Robert Poole, Marika Kutscher, Katrien Forier, Kevin Braeckmans, Wim Jiskoot] ABSTRACT: Purpose: To evaluate the potential of fluorescence single particle tracking (fSPT) for the characterization of submicron protein aggregates in human serum, plasma and formulations containing human serum albumin (HSA). Methods: A monoclonal IgG was covalently labeled with a fluorescent dye and cross-linked with glutaraldehyde. IgG aggregates and fluorescent beads of 0.1μm (control) were diluted in buffer, serum and plasma, and their size distributions were analyzed by fSPT and nanoparticle tracking analysis (NTA). In a separate experiment, IgG and HSA, fluorescently labeled with different dyes, were mixed and subjected to heat stress. The stressed sample was analyzed by fSPT using a dual color mode and by NTA. Results: The accuracy and precision of fSPT proved to be comparable to NTA. fSPT was able to successfully measure all the samples in buffer, serum and plasma. The average size of the cross-linked protein aggregates showed a slight increase in biological fluids. Moreover, fSPT analysis showed that a significant proportion of the aggregates formed by subjecting an IgG/HSA mixture to heat stress were composed of both proteins. Conclusion: fSPT is a powerful technique for the characterization of submicron protein aggregates in biological fluids and complex formulations. The Author(s), 2011 fluorescence single particle tracking nationallicence HSA nationallicence IgG nationallicence protein aggregates nationallicence serum and plasma nationallicence Filipe Vasco Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut Poole Robert Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut Kutscher Marika Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut Forier Katrien Biophotonic Imaging Group Laboratory of General Biochemistry & Physical Pharmacy Faculty of Pharmacy,, Ghent University, Harelbekestraat 72, 9000, Ghent, Belgium aut Braeckmans Kevin Biophotonic Imaging Group Laboratory of General Biochemistry & Physical Pharmacy Faculty of Pharmacy,, Ghent University, Harelbekestraat 72, 9000, Ghent, Belgium aut Jiskoot Wim Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/5(2011-05-01), 1112-1120 0724-8741 28:5<1112 2011 28 11095 https://doi.org/10.1007/s11095-011-0374-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-011-0374-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Filipe Vasco Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut NATIONALLICENCE 700 1- Poole Robert Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut NATIONALLICENCE 700 1- Kutscher Marika Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut NATIONALLICENCE 700 1- Forier Katrien Biophotonic Imaging Group Laboratory of General Biochemistry & Physical Pharmacy Faculty of Pharmacy,, Ghent University, Harelbekestraat 72, 9000, Ghent, Belgium aut NATIONALLICENCE 700 1- Braeckmans Kevin Biophotonic Imaging Group Laboratory of General Biochemistry & Physical Pharmacy Faculty of Pharmacy,, Ghent University, Harelbekestraat 72, 9000, Ghent, Belgium aut NATIONALLICENCE 700 1- Jiskoot Wim Division of Drug Delivery Technology Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/5(2011-05-01), 1112-1120 0724-8741 28:5<1112 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer