Production of cells and viral vectors
AssigneeBiocell Innovations Pte., Ltd.
InventorsPaula Lam, Mobashar Hussain Urf Turabe Fazil
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Abstract
Disclosed herein are cell culture methods, methods of producing a cell for use in adoptive cell therapy, methods of transducing an immune cell with a viral vector, and uses of products derived from the methods e.g. medical uses and therapeutic applications.
Claims34
A cell culture method, the method comprising adherent culture of cells in a culture vessel, wherein the cells are cultured in culture media that is a fetal bovine serum (FBS)-free media and the culture vessel contains a polypeptide comprising an amino acid sequence having at least 70% sequence identity to one of SEQ ID NO:3, SEQ ID NO:2, or SEQ ID NO:1, or a polypeptide comprising an amino acid sequence having at least 70% sequence identity to a fragment of SEQ ID NO:3, SEQ ID NO:2, or SEQ ID NO:1.
The method of any previous claim, wherein the cell is a mammalian or insect cell.
The method of any previous claim, wherein the polypeptide has at least 75% sequence identity to one of SEQ ID NO:3, SEQ ID NO:2, SEQ ID NO:1, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, and/or SEQ ID NO:23, preferably wherein the polypeptide has one of at least 80%, 85%, 90%, 95%, or 99% sequence identity to one of SEQ ID NO:3, SEQ ID NO:2, SEQ ID NO:1, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, and/or SEQ ID NO:23.
The method of any previous claim, wherein the polypeptide is present in the culture media.
The method of any previous claim, wherein the polypeptide is provided in the form of a liquid solution.
The method of any previous claim, wherein the polypeptide is coated on a surface of the culture vessel.
The method of any previous claim, wherein the culture vessel is a culture plate, a culture dish, a culture flask, or a bioreactor.
The method of any previous claim, wherein the polypeptide is provided as an additive to FBS-free media.
The method of any previous claim, wherein the culture media does not contain animal serum.
The method of any previous claim, wherein the cells have been transfected with a packaging helper plasmid.
The method of any previous claim, wherein the cells have been transfected with a plasmid encoding a gene of interest.
A method of producing a viral vector, the method comprising culturing cells according to the method of any previous claim, the method further comprising the step of collecting a virus, a viral vector, and/or a viral particle from the culture.
A method of producing a cell for adoptive cell therapy, the method comprising contacting a cell with a polypeptide comprising an amino acid sequence having at least 70% sequence identity to one of SEQ ID NO:3, SEQ ID NO:2 or SEQ ID NO:1, or a polypeptide comprising an amino acid sequence having at least 70% sequence identity to a fragment of SEQ ID NO:3, SEQ ID NO:2, or SEQ ID NO:1, and a viral vector encoding a gene of interest.
A method of transducing an immune cell with a viral vector encoding a gene of interest, the method comprising contacting immune cells with a viral vector encoding the gene of interest, and a polypeptide having at least 70% sequence identity to one of SEQ ID NO:3, SEQ ID NO:2 or SEQ ID NO:1, under conditions suitable for transduction of the cells with said viral vector to generate a transduced cell for adoptive cell therapy.
Dependencies are inferred from the claim text.