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Fluorescence single particle tracking for the characterization of submicron protein aggregates in biological fluids and complex formulations

Vasco Filipe, Robert Poole, Marika Kutscher, Katrien Forier UGent, Kevin Braeckmans UGent and Wim Jiskoot (2011) PHARMACEUTICAL RESEARCH. 28(5). p.1112-1120
abstract
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). A monoclonal IgG was covalently labeled with a fluorescent dye and cross-linked with glutaraldehyde. IgG aggregates and fluorescent beads of 0.1 mu m (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. 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. fSPT is a powerful technique for the characterization of submicron protein aggregates in biological fluids and complex formulations.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
ANTIBODY, QUANTITATION, IMMUNOGENICITY, NANOPARTICLES, serum and plasma, protein aggregates, IgG, fluorescence single particle tracking, HSA, COLLAGEN, ELISA, SERUM
journal title
PHARMACEUTICAL RESEARCH
Pharm. Res.
volume
28
issue
5
pages
1112 - 1120
Web of Science type
Article
Web of Science id
000289302000016
JCR category
PHARMACOLOGY & PHARMACY
JCR impact factor
4.093 (2011)
JCR rank
42/259 (2011)
JCR quartile
1 (2011)
ISSN
0724-8741
DOI
10.1007/s11095-011-0374-0
project
Center for nano- and biophotonics (NB-Photonics)
language
English
UGent publication?
yes
classification
A1
copyright statement
I have retained and own the full copyright for this publication
id
2128629
handle
http://hdl.handle.net/1854/LU-2128629
date created
2012-06-01 11:35:27
date last changed
2013-02-27 09:02:05
@article{2128629,
  abstract     = {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). 
A monoclonal IgG was covalently labeled with a fluorescent dye and cross-linked with glutaraldehyde. IgG aggregates and fluorescent beads of 0.1 mu m (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. 
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. 
fSPT is a powerful technique for the characterization of submicron protein aggregates in biological fluids and complex formulations.},
  author       = {Filipe, Vasco and Poole, Robert and Kutscher, Marika and Forier, Katrien and Braeckmans, Kevin and Jiskoot, Wim},
  issn         = {0724-8741},
  journal      = {PHARMACEUTICAL RESEARCH},
  keyword      = {ANTIBODY,QUANTITATION,IMMUNOGENICITY,NANOPARTICLES,serum and plasma,protein aggregates,IgG,fluorescence single particle tracking,HSA,COLLAGEN,ELISA,SERUM},
  language     = {eng},
  number       = {5},
  pages        = {1112--1120},
  title        = {Fluorescence single particle tracking for the characterization of submicron protein aggregates in biological fluids and complex formulations},
  url          = {http://dx.doi.org/10.1007/s11095-011-0374-0},
  volume       = {28},
  year         = {2011},
}

Chicago
Filipe, Vasco, Robert Poole, Marika Kutscher, Katrien Forier, Kevin Braeckmans, and Wim Jiskoot. 2011. “Fluorescence Single Particle Tracking for the Characterization of Submicron Protein Aggregates in Biological Fluids and Complex Formulations.” Pharmaceutical Research 28 (5): 1112–1120.
APA
Filipe, V., Poole, R., Kutscher, M., Forier, K., Braeckmans, K., & Jiskoot, W. (2011). Fluorescence single particle tracking for the characterization of submicron protein aggregates in biological fluids and complex formulations. PHARMACEUTICAL RESEARCH, 28(5), 1112–1120.
Vancouver
1.
Filipe V, Poole R, Kutscher M, Forier K, Braeckmans K, Jiskoot W. Fluorescence single particle tracking for the characterization of submicron protein aggregates in biological fluids and complex formulations. PHARMACEUTICAL RESEARCH. 2011;28(5):1112–20.
MLA
Filipe, Vasco, Robert Poole, Marika Kutscher, et al. “Fluorescence Single Particle Tracking for the Characterization of Submicron Protein Aggregates in Biological Fluids and Complex Formulations.” PHARMACEUTICAL RESEARCH 28.5 (2011): 1112–1120. Print.