Journal article
Expression, purification, and characterization of recombinant cyanovirin-N for vaginal anti-HIV microbicide development
Protein expression and purification, v 39(2), pp 229-236
2005
PMID: 15642474
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Cyanovirin-N (CV-N) is a prokaryotic protein under development as a topical anti-HIV microbicide, an urgent and necessary approach to prevent HIV transmission in at-risk populations worldwide. We have expressed recombinant CV-N as inclusion bodies in the cytoplasm of Escherichia coli. A purification scheme has been developed that exploits the physicochemical properties of this protein, in particular its stability in a harsh inclusion body purification scheme. Under the conditions developed, this system yields 140 mg of highly purified CV-N per liter of high-density cell culture, which represents a 14-fold increase over the best recombinant CV-N yield reported to date. This purification scheme results in monomeric CV-N as analyzed by SDS–PAGE, isoelectric focusing, and reverse phase- and size exclusion-HPLC. This recombinantly expressed and refolded CV-N binds to gp120 with nanomolar affinity and retains its potent anti-HIV activities in cell-based assays. The expression and purification system described herein provides a better means for the mass production of CV-N for further microbicide development.
Metrics
Details
- Title
- Expression, purification, and characterization of recombinant cyanovirin-N for vaginal anti-HIV microbicide development
- Creators
- Diana M. Colleluori - Biosyn, Incorporated, 1800 Byberry Road, Building 13, Huntingdon Valley, PA 19006, USADeborah Tien - Biosyn, Incorporated, 1800 Byberry Road, Building 13, Huntingdon Valley, PA 19006, USAFeirong Kang - Biosyn, Incorporated, 1800 Byberry Road, Building 13, Huntingdon Valley, PA 19006, USATara Pagliei - Biosyn, Incorporated, 1800 Byberry Road, Building 13, Huntingdon Valley, PA 19006, USARyan Kuss - Biosyn, Incorporated, 1800 Byberry Road, Building 13, Huntingdon Valley, PA 19006, USATimothy McCormick - Biosyn, Incorporated, 1800 Byberry Road, Building 13, Huntingdon Valley, PA 19006, USAKaren Watson - ImQuest BioSciences (United States)Karyn McFadden - Drexel UniversityIrwin Chaiken - Drexel UniversityRobert W. Buckheit - ImQuest BioSciences (United States)Joseph W. Romano - Biosyn, Incorporated, 1800 Byberry Road, Building 13, Huntingdon Valley, PA 19006, USA
- Publication Details
- Protein expression and purification, v 39(2), pp 229-236
- Publisher
- Elsevier
- Number of pages
- 8
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology
- Web of Science ID
- WOS:000226437000013
- Scopus ID
- 2-s2.0-19944427993
- Other Identifier
- 991019169681504721
UN Sustainable Development Goals (SDGs)
This publication has contributed to the advancement of the following goals:
InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Biochemical Research Methods
- Biochemistry & Molecular Biology
- Biotechnology & Applied Microbiology