Assembly of Human Papillomavirus Type-16 Virus-Like Particles: Multifactorial Study of Assembly and Competing Aggregation

Hanslip, J., Zaccai, R., Middelberg, A. P. J. and Falconer, R. (2006) Assembly of Human Papillomavirus Type-16 Virus-Like Particles: Multifactorial Study of Assembly and Competing Aggregation. Biotechnology Progress, 22 2: 554-560.


Author Hanslip, J.
Zaccai, R.
Middelberg, A. P. J.
Falconer, R.
Title Assembly of Human Papillomavirus Type-16 Virus-Like Particles: Multifactorial Study of Assembly and Competing Aggregation
Journal name Biotechnology Progress   Check publisher's open access policy
ISSN 8756-7938
8756-7938
Publication date 2006
Sub-type Article (original research)
DOI 10.1021/bp0502781
Volume 22
Issue 2
Start page 554
End page 560
Total pages 7
Editor Jerome S Schultz
Place of publication Washington
Publisher American Chemical Society
Collection year 2006
Language eng
Subject C1
290699 Chemical Engineering not elsewhere classified
291804 Nanotechnology
299999 Engineering and Technology not elsewhere classified
670499 Other
670799 Other
670199 Processed food products and beverages not elsewhere classified
100302 Bioprocessing, Bioproduction and Bioproducts
Abstract Pentameric capsomeres of human papillomavirus capsid protein L1 expressed in Escherichia coli self-assemble into virus-like particles (VLPs) in vitro. A multifactorial experimental design was used to explore a wide range of solution conditions to optimize the assembly process. The degree of assembly was measured using an enzyme-linked immunosorbent assay, and a high-throughput turbidity assay was developed to monitor competing aggregation. The presence of zinc ions in the assembly buffer greatly increased the incidence of aggregation and had to be excluded from the experiment for meaningful analysis. Assembly of VLPs was optimal at a pH of about 6.5, calcium and sodium ions had no measurable effect, and dithiothreitol and glutathione inhibited assembly. Tryptophan fluorescence spectroscopy demonstrated that an increase in urea concentration reduced the rate of VLP formation but had no effect on the final concentration of assembled VLPs. This study demonstrates the use of the hanging-drop vapor-diffusion crystallization method to screen for conditions that promote aggregation and the use of tryptophan fluorescence spectroscopy for real-time monitoring of the assembly process.
Keyword Food Science & Technology
In-vitro
Cervical-cancer
Capsid Protein
Young-women
L1
Fluorescence
Worldwide
Efficacy
Vaccine
Bovine
Q-Index Code C1

 
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Created: Wed, 15 Aug 2007, 10:37:29 EST