The present disclosure relates generally to the manufacturing of gene therapy products, and specifically to methods of purifying an enveloped virus from a cell culture fluid, comprising an endonuclease and/or anion exchange chromatography.

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 present invention relates to the industrialization of the production of recombinant lentiviral vectors in order to manufacture sufficient materials for therapeutic applications such as gene therapy and/or DNA vaccination, for use in clinical trials and/or commercial use.

The present disclosure relates to a combination of nucleic acids for the production of viral particles, said combination comprising or consisting of (a) a first nucleic acid encoding or being an inhibitory RNA; (b) at least one second nucleic acid comprising helper nucleic acids necessary for production of said viral particles, and/or encoding helper proteins necessary for said production; and (c) a third nucleic acid comprising a binding site for said inhibitory RNA. Furthermore, provided are methods of transfecting a cell with said combination, a production cell obtained by said transfecting or comprising the components of said combination and methods of producing viral particles.

The invention relates to methods of improving titer in transfection-based bioreactor culture production or transfection-based production systems using eukaryotic cells.

The present invention relates to a stock solution (RLP Stock Solution) of mammalian cell-endogenous retrovirus like particles (RLP), a method of preparing a RLP stock solution, a kit containing a RLP stock solution, and a method of quantifying the amount of RLP removed from a solution. An RLP stock solution will contain a high concentration of RLP and an extremely low amount of any therapeutic proteins of interest. In some instances, an RLP stock solution will contain a very low amount of natural mammalian host cell protein (HCP) and DNA. The method of preparing a RLP stock solution will consist of the production of RLP during fermentation or cell culturing and the subsequent purification of RLP from fermentation or cell culture solution. The kit will comprise at least two containers. One container comprised of RLP stock solution and one comprised of PCR primers or one or more antibodies. The method of quantifying RLP comprises the steps of adding RLP stock solution to an in-process solution containing a recombinant therapeutic of interest, processing the resulting solution through a bioprocess purification technique, and then quantifying the amount of RLP removed.

The invention relates to the preparation of a solution of polymer/nucleic acid complexes, and the use of such a solution in methods for the transfection of cells.

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 disclosure provides methods to chemically and photochemically initiate cell membrane blebbing to induce cell vesicle production, modifications thereof, and uses thereof, including for drug delivery, gene therapy, cell-free cell therapy, and molecular therapy.

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.