Method of producing recombinant biological products
AssigneeDublin City University
InventorsPatrick Gammell, Niall Barron, Martin Clynes
Life of this patent
Estimating the term…
Abstract
A method of producing a recombinant biological product, which method employs a mammalian producer cell culture, comprises the steps of generating a biomass of mammalian producer during an initial phase of cell culture, and causing an increase in a level of one or more of the miRNA molecules of Table 1 within the mammalian producer cells once a desired concentration of mammalian producer cells has been achieved. The method may also comprise the step of increasing a level of an inhibitor of one or more of the miRNA molecules of Table 1 within the mammalian producer cells at the start of or during an initial phase of culture.
Claims17
A method of producing a recombinant biological product comprising:
- (a) generating a biomass of Chinese Hamster Ovary (CHO) cells that produce said recombinant biological product at the start of or during an initial phase of cell culture;
- (b) causing an increase or decrease in a level of one or more miRNA molecules comprising a sequence selected from the group consisting of SEQ ID. NOs.:
- 9, 18, 21, and orthologous sequences thereof within the CHO cells,
- wherein said causing an increase comprises (i) transiently transfecting one or more of the miRNA molecules or precursors thereof within the CHO cells, (ii) transiently transfecting an expression vector comprising a nucleic acid sequence coding for one or more miRNA molecules or precursors thereof within the CHO cells, or (iii) stably transfecting a nucleic acid sequence coding for one or more miRNA molecules or precursors thereof into the CHO cell genome, and
- wherein said causing a decrease comprises increasing a level of an inhibitor of one or more the miRNA molecules;
- (c) reducing, arresting or increasing growth of the culture as a result of the increase or decrease in the level of the one or more miRNA molecules;
- and
- (d) producing the biological product from the culture.
A method of producing a recombinant biological product comprising:
- (a) generating a biomass of Chinese Hamster Ovary (CHO) cells that produce said recombinant biological product at the start of or during an initial phase of cell culture;
- (b) causing one or both of steps (b1) and (b2);
- and
- (b1) causing a decrease in a level of an miRNA molecule comprising a sequence selected from the group consisting of SEQ ID NO:
- 21 and an orthologous sequence thereof within the CHO cells at an initial stage of cell culture, thereby increasing growth of the cell culture as a result of the decrease in the level of the miRNA molecule;
- and
- (b2) causing an increase in a level of an miRNA molecule comprising a sequence selected from the group consisting of SEQ ID NO:
- 21 and an orthologous sequence thereof within the CHO cells at a growth arrest phase, thereby reducing or arresting growth of the culture as a result of the increase in the level of the miRNA molecule,
- wherein said causing a decrease comprises increasing a level of an inhibitor of the miRNA molecule, and
- wherein said causing an increase comprises (i) transiently transfecting the miRNA molecule or a precursor thereof within the CHO cells, (ii) transiently transfecting an expression vector comprising a nucleic acid sequence coding for the miRNA molecule or a precursor thereof within the CHO cells, or (iii) stably transfecting a nucleic acid sequence coding for the miRNA molecule or a precursor thereof into the CHO cell genome, and;
- (c) producing the biological product from the culture.
Dependencies are inferred from the claim text.