Production of Soluble Human Gamma Interferon in the  Escherichia coli  Expression System with a Decrease in Cultivation Temperature

E. A. Volosnikova; Волосникова Е. А.; T. I. Esina; Есина Т. И.; D. N. Shcherbakov; Щербаков Д. Н.; N. V. Volkova; Волкова Н. В.; Ya. S. Gogina; Гогина Я. С.; T. A. Tereshchenko; Терещенко Т. А.; E. D. Danilenko; Даниленко Е. Д.

doi:10.31857/S0555109923020174

Production of Soluble Human Gamma Interferon in the Escherichia coli Expression System with a Decrease in Cultivation Temperature

Autores: Volosnikova E.A.¹, Esina T.I.¹, Shcherbakov D.N.¹, Volkova N.V.¹, Gogina Y.S.¹, Tereshchenko T.A.¹, Danilenko E.D.¹
Afiliações:
1. State Research Center of Virology and Biotechnology “Vector”
Edição: Volume 59, Nº 2 (2023)
Páginas: 167-173
Seção: Articles
URL: https://cardiosomatics.ru/0555-1099/article/view/674632
DOI: https://doi.org/10.31857/S0555109923020174
EDN: https://elibrary.ru/LVLPXU
ID: 674632

Citar

Texto integral

Acesso aberto
Acesso é fechado

Acesso está concedido
Acesso é fechado

Acesso é pago ou somente para assinantes

Resumo
Sobre autores
Bibliografia
Arquivos suplementares
Estatísticas

Resumo

A recombinant strain producing human gamma interferon (IFN-γ) E. coli BL 21/pET-IFN-γ was constructed, providing a high level of its expression. A method has been developed for obtaining a soluble form of recombinant IFN-γ, consisting of the processes of producing a biomass of a producer strain containing a target protein in an amount of 32–37% of the total content of cellular proteins, protein isolation and purification. The purification process included the stages of disintegration, clarification of the cell lysate, chromatographic purification and dialysis. The developed method makes it possible to obtain from 1 g of wet biomass up to 5 mg of the drug with a purity of at least 95% and high specific (antiviral) activity.

Palavras-chave

human recombinant gamma interferon, producer strain, cultivation, chromatography, specific activity

Bibliografia

Pandey R., Prabhu A.A., Dasu V.V. // Separation Science and Technology. 2018. V. 53. № 3. P. 487–495. https://doi.org/10.1080/01496395.2017.1395463
Razaghi A., Owens L., Heimann K. // J. Biotechnol. 2016. V. 240. P. 48–60. https://doi.org/10.1016/j.jbiotec.2016.10.022
Khalilzadeh R., Shojaosadati S.A., Bahrami A., Maghsoudi N. // Biotechnology Letters. 2003. V. 25. № 23. P. 1989–1992. https://doi.org/10.1023/b:bile.0000004390.98648.25
Rosano G.L., Ceccarelli E.A. // Front. Microbiol. 2014. V. 5. P. 172. https://doi.org/10.3389/fmicb.2014.00172
Malhotra A. // Methods in Enzymol. 2009. V. 463. P. 239–258. https://doi.org/10.1016/S0076-6879(09)63016-0
Leibly D.J., Nguyen T.N., Kao L.T., Hewitt S.N., Barrett L.K., Van Voorhis W.C. // PlOS ONE. 2012. V. 7. №. 12. P. e52482. https://doi.org/10.1371/journal.pone.0052482
Tileva M., Krachmarova E., Ivanov I., Maskos K., Nacheva G. // Protein Expr. Purif. 2016. V. 117. P. 26–34. https://doi.org/10.1016/j.pep.2015.09.022
Государственная фармакопея РФ XIV изд. Т.2. ОФС.1.7.2.0002.15 Биологические методы испытания препаратов интерферона с использованием культур клеток. С. 2740–2749. https://femb.ru/record/pharmacopea14
Хайруллин Р.Ф. Экспрессия рекомбинантных белков в E. coli: учебн. пособие. / Ред. Р.Ф. Хайруллин, Р.Г. Киямова, А.А. Ризванов. Казань: Изд-во Казанского ун-та, 2018. 142 с.
Закабунин А.И., Барановская Г.А., Пустошилова Н.М., Майстренко В.Ф., Гаврюченкова Л.П., Громова О.А. Способ получения рекомбинантного интерферона – гамма человека. Патент РФ. 1999. № 2132386.