8/31/2023 0 Comments Capto q impress pore size![]() In addition, the existing chromatography purification methods also have the disadvantages of complicated steps, low yield and low purity, etc, and are hard to meet the requirements of industrialized large-scale preparation and GMP grade production. The prepared lentivirus vector may have high residues of endotoxin, BSA, HCP or nucleic acid, and cannot be administrated directly into human body. Although this method is simple, it cannot be industrially used for large-scale production. The method used in conventional laboratories to obtain lentivirus is ultracentrifugation. The mature lentiviral particles are secreted by 293T cells into the culture supernatant, and are obtained by ultracentrifugation or chromatography methods. Mature lentiviral particles are produced by co-transfecting 293T cells with multiple vectors and packaging in cells. The current recombinant lentivirus vector is genetically modified to only retain the packaging signal and target gene transcripts in the lentiviral genome, while the structural genes such as reverse transcriptase, envelope protein VSVG and gag-pol are separated in 2 or 3 vectors, and disease-causing genes are removed simultaneously. Recombinant lentivirus vector has become the first transgenic vectors for CART cells and gene therapy due to its high biological titer and low immunogenicity in vivo and in vitro. Different from the general retroviral vector, it has the ability to infect both dividing cells and non-dividing cells. Recombinant lentivirus vector is a gene therapy vector developed based on HIV-1 (Human Immunodeficiency Type I Virus). The transduction efficiency of viral vectors is much higher than that of non-viral vectors, and is particularly suitable for infecting target cells that are difficult to be infected, such as lymphocytes. ![]() Common viral vectors comprise recombinant retrovirus (rRV), recombinant lentivirus (rLV), recombinant adenovirus (rAd), and recombinant adeno-associated virus (rAAV), etc. Viral vectors encapsulate exogenous genes with the shell of natural viruses, and use the infectivity of virus against host cells to introduce exogenous genes into cells. Non-viral gene delivery vectors are relatively safe and stable, but their transfection efficiency is usually low. Commonly used non-viral vectors comprise liposome, dendrimer, non-natural cationic polymer and natural polysaccharide, etc. ![]() Gene therapy refers to the introduction of exogenous therapeutic genes into target cells to correct or compensate diseases caused by gene defects and abnormalities, or refers to using products expressed by exogenous genes to act on disease targets, thereby achieving the purpose of treatment.Įxogenous genes can be transduced or delivered through viral or non-viral vectors. The present invention relates to the field of biotechnology and specifically, relates to a method for large-scale preparation of purified preparation of recombinant lentiviral vector at GMP grade. 28, 2018, and which incorporates by reference those PCT and Chinese applications in their entireties. 28, 2019, which claims priority from Chinese Patent Application Serial No. § 371 of Patent Cooperation Treaty Application No. ![]() national phase application under 35 U.S.C.
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