Provided are a CYBB lentiviral vector, a lentiviral vector-transduced stem cell, a preparation method and application thereof. The lentiviral vector includes a hEF 1α promoter and CYBB that are organized in tandem. The lentiviral vector carries the CYBB gene which under the initiation of the hEF 1α promoter, and expresses the carried CYBB gene in differentiated or undifferentiated stem cells. Stem cells serve as a delivery vector.

The present invention relates to the development of a method for the manufacturing of LVV vectors in a packed bed bioreactor by multi-plasmid transient DNA transfection. More particularly, the present invention discloses and claims a process in which steps applied in perfusion or in batch mode are combined to obtain an effective process. Moreover, the invention identifies an effective range of total DNA to be successfully used in the step of transient transfection in a packed bed bioreactor for the manufacturing of lentiviral vectors.

Methods of using viruses labeled with alkyne-modified biomolecules, such as fatty acids, carbohydrates and lipids, to treat a plant, an insect or an animal infected with a virus or to increase the infectivity of a virus, such as the human immunodeficiency virus, are provided. Also provided are methods of labeling a virus, such as human immunodeficiency virus, with an alkyne-modified biomolecule, such as a fatty acid, a carbohydrate, or an isoprenoid lipid. The viruses labeled with alkyne-modified biomolecules may be combined with a pharmaceutically acceptable excipient to produce a pharmaceutical composition, optionally containing another anti-viral agent and/or a delivery agent, such as a liposome.

Disclosed is an application of methioninase gene therapy in the treatment of a malignant tumor. In the gene therapy, a virus is used as a vector to insert an exogenous methioninase gene to constitute a methioninase expression system inside a tumor, thus providing an endogenous mechanism for further consumption of methionine. In vitro cytology experiments indicate that the methioninase gene therapy significantly reduces the level of intracellular methionine, effectively inhibits the proliferation of tumor cells, and can be used in an application in preparing a drug for the targeted treatment of a malignant tumor.

The present disclosure provides an automated method of producing viral vectors, utilizing engineered viral vector-producing cell lines within a fully-enclosed cell engineering system. Exemplary viral vectors that can be produced include lentivirus vectors, adeno-associated virus vectors, baculovirus vectors and retrovirus vectors.

Stabilized trimers of a clade A strain and a clade C strain of HIV-1 are provided. Broadly neutralizing antisera against HIV-1, methods of making broadly neutralizing antisera against HIV-1, broadly neutralizing vaccines against HIV-1, as well as methods of treating subjects infected with HIV, preventing HIV infection, and inhibiting HIV-mediated activities are also provided.

Provided is a method for large-scale production of lentivirus by using GMP-level serum-free suspension cells. Said method comprises the following steps: (a) providing a seed solution of packaged cells; (b) inoculating the seed solution in a first culture solution; (c) carrying out subculture of the packaged cells; (d) starting a liquid change operation when a liquid change trigger condition is met; (e) repeating steps (c) and (d) 1, 2 or 3 times; (f) starting a transfection operation when a transfection trigger condition is met; (g) optionally performing liquid change after transfection; (h) cultivating the transfected packaged cells; (i) starting harvesting and liquid change operations when a liquid change trigger condition is met; (j) repeating steps (h) and (i) 1, 2 or 3 times; (k) combining each of the recovered liquids; and (1) performing a purifying treatment. The culture solution used in each step is a serum-free cell culture solution.

The present invention provides a method and a reagent for increasing lentiviral vector production in 293T cells. When the 293T cells are cotransfected with a packaging mix comprising a plurality of plasmids that comprise genes encoding proteins essential for lentiviral particle formation and a plasmid that comprises a gene transcribed into RNA to be incorporated into lentiviral particles containing a transgene to be expressed in target cells according to the method for producing a lentiviral vector, HTLV-1 Tax, HIV-1 Tat, or NF-κB RelA are coexpressed in the 293T cells, so as to increase the amount of lentiviral vector production.

The present invention provides methods of making engineered viral proteins and protein complexes that are useful as vaccine immunogens, engineered viral proteins and protein complexes made using such methods, and pharmaceutical compositions comprising such engineered viral proteins and protein complexes. Such engineered viral proteins and protein complexes may comprise one or more cross-links that stabilize the conformation of an antibody epitope, such as a quaternary neutralizing antibody, and may exhibit an enhanced ability to elicit a protective immune response when administered to a subject as a component of a vaccine.

In accordance with the present invention, a method for increasing the yield of rLV vector particles comprising a trans gene encoding a therapeutic protein or fragment thereof is disclosed. In one approach, cells are transfected with plasmids encoding the necessary components for rLV production using a calcium chloride transfection mix at pH 7.1 wherein the calcium chloride and plasmids form a complex which is added to the cells at a constant speed. The cells are then incubated for a suitable time period wherein virus particle media is collected at least twice during the incubation period and stored in a cold storage unit, thereby reducing virus inactivation.