Influence of Glucose Concentration in Culture Medium on Recombinant gp 120 HIV-1 Production

Contingent screening for specific serological markers of infection ranks high in HIV prophylaxis. Nowadays the most common serological screening practice of HIV is contingent testing to detect serum antibodies against the HIV using an enzyme linked immunosorbent assay (ELISA). Recombinant analogues of HIV-1 envelope protein gp 120 are widely used in all commercial ELISA test kits for HIV diagnostics. In this paper we study the production of rgp 120 in E.coli host strain. The research strategies focus on improving the cultivation techniques and manipulating the bacteria physiology. Two fed batch strategies are applied – with continuous and exponential feeding. Exponential feeding is used so that the cells can be grown at the desired growth rate (μ) by programming the bioreactor software. By providing proper nutrient and operation conditions, this strategy allows us reaching a high cell concentration and achieving high yields of recombinant product.

Publication year: 
С. 61—65. Іл. 2. Табл. 1. Бібліогр.: 8 назв.

1. J. Shiloacha and R. Fass, “Growing E.coli to high cell density — a historical perspective on method development”, Biotechnology Advances, vol. 23, pp. 345—357, 2005.
2. Y. Morikawa et al., “HIV-l envelope protein gp 120 expression by secretion in E. coli: assessment of CD4 binding and use in epitope mapping”, J. of Virological Method, vol. 29, pp.105—114, 1990.
3. D.J. Korz et al., “Simple fed-batch technique for high cell density cultivation of Escherichia coli”, J. of Biotechnology, vol. 39, pp. 59—65, 1995.
4. J. Aulicino and M. Hermida, “Developing an automatically controlled feeding process in an E.coli fermentation process for recombinant protein production”, The Science& Business of Biopharmaceuticals, vol. 6, pp. 2—7, 2010.
5. U. Laemmli, “Cleavage of structural proteins during the assembly of the head of bacteriophage T4”, Nature, vol. 227, pp. 680—685, 1970.
6. pET System Manual, Novagen, 2003, 68 p.
7. Jong Hyun Choia et al., “Production of recombinant proteins by high cell density culture of Escherichia coli”, Chem. Eng. Sci., vol. 61, pp. 876—885, 2006.
8. K. Hellmuth et al., “Effect of growth rate on stability and gene expression of recombinant plasmids during continuous and high cell density cultivation of Escherichia coli TG1”, J. of Biotechnology, vol. 32, pp. 289—298, 1994.

2012-3-10.pdf258.02 KB

Тематичні розділи журналу