Provided are a human PD-1 knockdown siRNA, a recombinant expression CAR-T vector, a preparation method thereof, and an application of the same. A PD-1 knockdown siRNA expression cassette and an siRNA expression product thereof can be applied to a CAR-T therapy of multiple myeloma (MM) for eliminating or alleviating a tumor immune escape mechanism, and in the suppression of an immune escape mechanism in a CAR-T therapy of a tumor, such as pancreatic cancer, brain glioma, and myeloma.

We have modified a commercially-available adherent cell culture bioreactor, developing a new sampling manifold configuration and new way of taking samples to reduce contamination risk.

The present invention relates to a producer cell which expresses a tagging protein at the cell surface, such that retroviral vectors produced by the cell are tagged with the tagging protein, wherein the tagging protein comprises: i) a binding domain which binds to a capture moiety ii) a spacer; and iii) a membrane targeting domain such that, when incorporated a retroviral vector, the tagging protein facilitates purification of the retroviral vector from cellular supernatant via binding of the tagging protein to the capture moiety. The present invention also relates to a retroviral vector comprising such a producer cell-derived tagging protein.

This disclosure relates to the use of size exclusion chromatography and/or size exclusion chromatography with multi-angle light scattering technology to characterize viral particles such as adeno-associated virus and lentivirus particles. The disclosed methods are also useful for estimating the titer of viral particles, determining the integrity of the viral particles and estimating the amount of DNA encapsidated in the viral particle.

Viral vector production processes and methods of purifying a viral vector from a host cell are provided herein.

The present invention contemplates mRNA, episomal and retroviral genomic gene therapy based short-term, intermediate or long-term vaccine, immunization, immune protection or cancer—that can also be administered as a retroviral genomic gene therapy both in vivo and ex vivo—method to provide epithelial and hematological protection to humans to protect against cancer especially carcinomas, pandemic and non-pandemic viruses, bacterial infections, allergens or the cause of allergic reactions, systemic pathological conditions, cancer and anti-biowarfare agents (e.g. natural and unnatural viruses and toxins) where mucosal immunity and for some diseases hematological immunity is achieved through mRNA, episomal or genomic integrated lentiviral and gammaretroviral vector expression of dimeric immunoglobulin A1 (dIgA1), dimeric immunoglobulin A2 (dIgA2) and engineered variants. Additionally, in some embodiments a method to agglutinate cancers including carcinomas and hematological cancers to prevent metastasis with polymeric immunoglobulin A and dimeric immunoglobulin A and engineered variants. The present invention provides methods, immunoglobulin compositions and vector constructs to express potent immunoglobulins that are derived from human blood of a human currently infected with, affected by, exposed to or recovered from any of a wide range of allergens or the cause of allergic reactions, pathogens (including, viruses, virus mutants, bacterial infections and fungi) and systemic pathological ailments (including cancer and other disorders), developed from phage display technology or mice or other non-human vertebrates with engineered immune systems or humanized immune systems, transgenic mice or chimeric antibodies a fusion of non-human vertebrates (e.g. mouse or rabbit), mouse antibody V-regions, human antibodies. The immunoglobulin compositions include the heavy chain variable, diversity and joining (VDJ or Variable Heavy Region genes) segment immunoglobulin DNA and/or polypeptide sequence from humans identified to have therapeutically relevant affinity immunoglobulins against the antigen, protein or proteins of interest and either to use the exact immunoglobulin heavy chain and light chain polypeptide sequences identified from the B-cell that produced them or to modify or engineer some of the immunoglobulin heavy chain and light chain constant domains to modulate effector functions. Although, ideally there are no changes made to the immunoglobulins light and heavy chains as identified from the B-cell that produced them. Modifications may occur at the Hinge region, Constant Heavy 2 (C.sub.H2) domain and Constant Heavy 3 (C.sub.H3) domain for the immunoglobul

The present invention describes the development of a modified envelope glycoproteins to pseudo-type viruses of the retroviridae family. These are derived from Murine leukaemia virus amphotropic, Gibbon Ape leukaemia virus and feline endogenous virus envelopes.

The improved envelope glycoproteins contain, among other modifications, newly introduced alternative cleavable sequences.

The viral vectors pseudo-typed with these modified envelopes may be suitably employed for cargo delivery such as in gene and cell therapy applications, for the ex vivo and in vivo delivery of gene(s), protein(s), or molecule(s) of interest to a variety of target cells.

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.

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.

Provided for herein are fusogenic rhabdovirus glycoproteins and uses thereof, compositions comprising the same, and methods of using the same. Also provided for herein are pseudotyped viral particles comprising rhabdovirus glycoproteins as provided for herein and targeting moieties as provided for herein. Also provided are methods of generating and using the pseudotyped viral particles as provided for herein.