Lentivirus-Based System for Fast Expression and Purification of Recombinant Proteins and its Implementation for Production of Human CD44 Extracellular Part

Yulia Evgenyevna Kravchenko¹, Stepan Petrovich Chumakov¹, Elena Ivanovna Frolova^1,*, Yuriy Nikolaevich Lezhnin^1,2, Dmitry Sergeevich Kravchenko^1,2

¹Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, 117997, Russian Federation, Moscow, GSP-7, Ulitsa Miklukho-Maklaya, 16/10

²Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, Russia

DOI : http://dx.doi.org/10.13005/bbra/1884

ABSTRACT: Development of novel highly selective anticancer therapeutics usualy requires large amount of target protein that serves as a bait for selection. In many cases these proteins posess posttranslational modifications or features, that could not be replicated in procaryotic expression systems, thus requiring more laborous and expensive expression in mammalian cells. Transfection-based eucaryotic producents, on the other hand, are unstable and gradually lose production yields.  We developed novel lentivirus-based expression system for fast and simple production and purification of recombinant proteins in mammalian cells. Our expression system allows easy selection of producent cells with flow sorting, antibody-free assessment of secreted protein concentration in cellular media and fast two-stage purification of target protein devoid of any tag sequences. We implemented our expression system for fast production of highly efficient mammalian producent of CD44 receptor extracellular part and subsequent purification of properly folded and glycosylated protein

KEYWORDS: Lentiviral vectors; CD44; tagRFP; protein expression  

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