Lentivirus production

Viral vector production-Gene Transfer、Gene expression、Gene editing

Lentiviral vectors have been widely used in gene and cell therapy, and are the key material of CAR-T cells that have been popular in recent years. They are currently very effective tools in molecular and cellular biological experiments. As a type of retrovirus-derived vectors, Its transfection efficiency is high, and it can infect cells that are in the active phase of mitosis, slow division and the end of division. The lentiviral vector carries a large number of important genetic information into the host cell to obtain long-term and stable expression. In addition, the lentiviral vector does not express any HIV-I protein and has low immunogenicity. The modified lentiviral vector can accommodate about 10kb of foreign genes, so most of the cDNA can be cloned into the lentiviral vector. These advantages make lentiviral vector an effective tool for gene transfer in vivo and in vitro, and it has a wide range of application prospects.

Types Of Viral Vectors

2nd generation lentiviral vector

The second-generation three-plasmid expression system is to separate the cis-acting sequence structure and the sequence encoding the trans-acting protein required for packaging, reverse transcription and integration in the HIV-1 genome, and then clone them into three independent plasmids Among them, one is a packaging plasmid, which contains the CMV promoter, which controls the expression of all viral structural genes except env, and the second is an envelope plasmid, which contains the sequence of the vesicular stomatitis virus glycoprotein (VSV-G) gene, This gene is used to replace the env gene of the original virus. The product of this gene can increase the host range of the virus, and the other is a vector plasmid, which contains the sequence structure of the gene that researchers are interested in. However, this three-plasmid system still retains the accessory genes of HIV-1. This plasmid system is more likely to have an active virus due to accidents, so it is considered to have a lower safety factor, but because of its higher potency for virus production , More suitable for research use.

3rd generation lentiviral vector

The third-generation lentivirus system. Researchers have also made further improvements to the second-generation three-plasmid system. This is also a lentiviral vector system that is currently widely used in clinical practice. It removes all auxiliary sequences of the HIV virus. The original HIV 9 Three genes (gag, pol, rev) are kept in the constructed lentiviral vector, and the env gene is placed on a separate expression plasmid. In addition, the tat gene is removed, thus forming a four-plasmid system , Namely Gag/Pol, Rev, VSV-G, in addition, there is a vector that can place the target gene sequence. This combination drastically reduces the possibility of recombinant active virus.

Retain Biotech's technology

Retain Biotech has successfully developed a technology platform for the production of high-efficiency lentivirus and applied it to our existing CAR-T process. Different from common lentiviruses, it can effectively improve the production, concentration and transfection efficiency of lentivirus after optimization by our technology; Retain Biotech can provide two generations of virus packages and two generations of RUO and Pre-clinical. Compared with the general second-generation lentiviral technology, the third-generation lentiviral technology greatly improves the safety of genetically modified cells and reduces the risk of patients. In addition, we are also developing a semi-automated instrument production method, which can further increase the production of lentivirus, production efficiency and reduce human operations.

OEM Viral Vector Service

Research grade Lentivirus

Accept commissioned OEM to develop and produce 2nd generation and 3rd generation viral vectors

Pre-Clinical grade Lentivirus

Accept commissioned OEM to develop and produce 2nd generation and 3rd generation viral vectors

The advantages of Retain Biotech's viral vector production technology

High titer

Can provide >10⁷ TU/ml high titer lentivirus

High production volume

Can provide ≤ 1500ml high volume lentivirus

High quality

Can provide pass >5 QC test items lentivirus