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

Yulia Kravchenko, Stepan Chumakov, Elena Frolova, Yuriy Lezhnin, Dmitry Kravchenko
2015 Biosciences Biotechnology Research Asia  
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
more » ... ivirus-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.
doi:10.13005/bbra/1884 fatcat:eeib6xyqj5ekjmyzpimahrvu3e