Lentiviral vector (LV) formulations, and pharmaceutical compositions comprising such LV formulations, with improved stability and suitable for systemic administration are provided. Methods for treating disorders, especially blood disorders, using systemic administration of LV formulations are also provided.

Disclosed herein are viral vector production systems secreting nuclease for degradation of residual nucleic acid during viral vector production and methods of the same. Such a viral vector production system comprises a viral vector production cell comprising nucleic acid sequences encoding: 1) viral vector components; and 2) a nuclease, wherein the nuclease is expressed in the production cell and secreted in cell culture thereby degrading residual nucleic acid during viral vector production. Another such viral vector production system comprises 1) a viral vector production cell comprising nucleic acid sequences encoding viral vector components; and 2) a nuclease helper cell comprising a nucleic acid sequence encoding a nuclease, wherein the nuclease is expressed and secreted in co-culture of the production cell of 1) and the helper cell of 2), thereby degrading residual nucleic acid during viral vector production.

The present disclosure relates to methods for producing lentiviral vector-producing cells. Specifically the methods utilize two plasmids, rather than four, to provide the required packaging elements and transfer vector to a cell, allowing for the production of a large number of lentiviral producer cells, including suspension-based cells, and the production of high amounts of lentivirus. These methods allow for the production of cells that can be later induced to produce lentivirus, and can be tailored to include a specific gene of interest.

Provided is a method for large-scale preparation of a purified preparation of a recombinant lentiviral vector at the GMP grade. The method comprises: (a) providing raw material feed liquid to be purified that comprises recombinant viral vectors; (b) carrying out a microfiltration treatment on the feed liquid to obtain a microfiltered filtrate comprising the recombinant viral vectors; (c) optionally concentrating the filtrate to obtain a concentrated filtrate; (d) purifying the filtrate obtained in the previous step by means of chromatography to obtain a crude pure product comprising the recombinant viral vectors; and (e) subjecting the crude pure product obtained in the previous step to liquid exchange and elaborate purification to obtain the purified recombinant viral vectors.

Novel forward primer, reverse primer and poly-linker suitable for replication of nucleic acids in e.g., 293 cells.

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.

The invention provides homology directed repair (HDR) constructs for variant screening in cells comprising: a left and right homology arm, with either the left or right homology arm encoding a genomic edit to be incorporated at a target locus; and an excisable double selection cassette located within the left and right homology arms, the excisable double selection cassette comprising; a first selection marker; and a second selection marker; and wherein the first selection marker and the second selection marker are located between a first and second excision site. Also provided are homology directed repair (HDR) vectors comprising a construct as described herein, and methods for using such vectors.

The invention provides lentiviral preparations containing a sulfonic acid buffer, such as 1,4-piperazinediethanesulfonic acid (PIPES), 2-(N-morpholino)ethanesulfonic acid (MES), and 3-morpholinopropane-1-sulfonic acid (MOPS), a sodium citrate buffer, or a phosphate buffer. The invention additionally encompasses methods of lentiviral purification as well as methods of transducing human cells.

This invention relates to lentiviral gene transfer vectors pseudotyped with hemagglutinin-neuraminidase (HN) and fusion (F) proteins from a respiratory paramyxovirus, comprising a promoter and a transgene; and methods of making the same. The present invention also relates to the use of said vectors in gene therapy, particularly for the treatment of respiratory tract diseases such as Cystic Fibrosis (CF).

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.