Chapter title |
Production of Well-Characterized Virus-like Particles in an Escherichia coli-Based Expression Platform for Preclinical Vaccine Assessments.
|
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Chapter number | 29 |
Book title |
Vaccine Design
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Published in |
Methods in molecular biology, January 2016
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DOI | 10.1007/978-1-4939-3389-1_29 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3388-4, 978-1-4939-3389-1
|
Authors |
Newton Wahome Ph.D., Anne Cooper, Prem Thapa, Shyamal Choudhari, Fei P. Gao, David B. Volkin, C. Russell Middaugh, Newton Wahome |
Editors |
Sunil Thomas |
Abstract |
In this chapter we demonstrate a method to produce virus-like particles (VLPs) from Escherichia coli. Standard bacterial protocols are used for the cloning, transformation, and expression of the protein subunits. A two-step protein purification method is highlighted: one step based on separating soluble proteins with ion-exchange affinity chromatography and a second polishing step using size-exclusion columns to isolate VLP species. The ensuing VLPs can be characterized with a variety of biophysical techniques including ultraviolet (UV)-visible spectroscopy for protein quantification, dynamic light scattering for size distribution determination, and transmission electron microscopy to ascertain size and morphology. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 11 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Master | 4 | 36% |
Student > Bachelor | 1 | 9% |
Other | 1 | 9% |
Student > Ph. D. Student | 1 | 9% |
Researcher | 1 | 9% |
Other | 0 | 0% |
Unknown | 3 | 27% |
Readers by discipline | Count | As % |
---|---|---|
Immunology and Microbiology | 3 | 27% |
Chemical Engineering | 1 | 9% |
Veterinary Science and Veterinary Medicine | 1 | 9% |
Biochemistry, Genetics and Molecular Biology | 1 | 9% |
Engineering | 1 | 9% |
Other | 0 | 0% |
Unknown | 4 | 36% |