Effective Transduction of Brain Neurons with Lentiviral Vectors Purified by Ion-Exchange Chromatography

Abstract

The development of methods for purification viral vectors for gene therapy is one of the most important and urgent problems of modern biology and medicine. Recently, drugs that carry cerebral neurotrophic factors, such as BDNF, have become increasingly popular. However, viral drugs for gene therapy should meet certain requirements, including high titer and applicability for in vivo studies. At the same time, the creation of such vectors requires cost-effective, inexpensive and affordable methods for standard laboratories. This study compares various methods for purification of lentiviral vectors encoding the brain neurotrophic factor, BDNF. The highest titer (1.12 ∙ 109/mL) was obtained via PEG 6 000 precipitation followed by anion-exchange chromatography on two columns of sorbents containing quaternary ammonium groups. Abnormal aggregates of transduced neurons were detected after lentiviruses purified only by PEG precipitation were injected into the brain of a newborn rat. This fact confirms the necessity of the proposed additional chromatographic purification stage. lentivirus, BDNF, ion exchange chromatography, gene therapy, PEG This work was financially supported by budgetary funding project no. 0259-2019-0003-C-01.