DEVELOPMENT OF A MEDICINAL AGENT BASED ON THE PEPTIDE ANTAGONIST OF OREXIN RECEPTOR FOR SUPPRESSION OF ALCOHOL DEPENDENCE



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Abstract

The aims of the research were to synthesize gene, coding previously designed artificial protein antorex - the antagonist of orexins receptor OxR1, to explore it's expression in Escherichia coli cells and to develop the process of this protein purification from Escherichia coli cell lysates. Methods of chemical and enzymatic DNA synthesis, genetic engineering, selection and chromatography were used. The gene coding antorex protein was synthesized, the plasmid DNA рЕТ-antorex, containing this gene and Escherichia coli strain BL21(pЕT-аntorex) - the stable producer of recombinant protein antorex were obtained. Methods of Escherichia coli producer strain cultivation and antorex protein purification with the use of reverse-phase and ion-exchange chromatography was developed. Large-scale cultivation of producer strain was performed in 10L fermenter and the pilot lot of 480 mg of antorex protein with the purity of 97,5% was produced. The developed methods lead to production and purification of antorex artificial protein for conducting pre-clinical and clinical trials.

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About the authors

I V Duhovlinov

Institute of Experimental Medicine

N A Klimov

Institute of Experimental Medicine

E A Fedorova

Institute of Experimental Medicine

Yu V Zherebcova

Institute of Experimental Medicine

E A Chernyaeva

Institute of Experimental Medicine

A I Orlov

Institute of Experimental Medicine

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Copyright (c) 2017 Duhovlinov I.V., Klimov N.A., Fedorova E.A., Zherebcova Y.V., Chernyaeva E.A., Orlov A.I.

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