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Enhancement of cellular production through mechanotransduction

A technology of cell and cell adhesion, applied in stress-stimulated microbial growth methods, biochemical instruments, biochemical equipment and methods, etc.

Inactive Publication Date: 2012-05-23
ABBVIE INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Cell culture methods using mammalian cells, including CHO cells, for the production of biotherapeutics face many challenges, including compressed product development timelines, insufficient capacity, and limitations in cell line proliferation and productivity

Method used

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  • Enhancement of cellular production through mechanotransduction
  • Enhancement of cellular production through mechanotransduction

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Experimental program
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Embodiment

[0069] 1. Cell culture

[0070] CHO cells producing recombinant, glycosylated antibodies (eg, ABT-874) are used. ABT-874 producing floating CHO cells were adapted for growth in serum. Cells were cultured on cell culture plates under the nominal growth conditions used in fed-batch reactors.

[0071] 2 Generation of tensegrity forces on cells

[0072] Cells were introduced into the tensegrity model using two different techniques as described in sections 2.1 and 2.2 below. Cell-based assays were used to optimize and evaluate changes in mechanotransduction induced by these forces.

[0073] 2.1 Optical magnetic torsional cytometry

[0074] At various time points, cells are subjected to controlled mechanical shear stress applied directly to the cell surface. Shear stress (torque) is applied to the surface using membrane-bound ferromagnetic beads (1-6 μm in diameter) coated with anti-CHO antibody that adheres to the cell's cytoskeleton. The beads are magnetized in one direction...

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Abstract

Disclosed herein are methods of modulating protein production via the application of tensegrity forces on cells and cell cultures. The methods of the invention increase production of protein from cells and cell culture. The tensegrity forces can be stress that is applied to the cells, and can include one or more of the following; mechanical stress, shear stress, stretch effects, and pressure induced stress.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Application Serial No. 61 / 225,694, filed July 15, 2009, which is hereby incorporated by reference in its entirety. Background of the invention [0003] Chinese hamster ovary (CHO) cells are the most commonly used mammalian host cell line for the production of recombinant proteins in the biopharmaceutical industry. Cell culture methods using mammalian cells, including CHO cells, for the production of biotherapeutics face numerous challenges, including compressed product development timelines, insufficient capacity, and limitations in cell line proliferation and productivity. [0004] The present invention addresses these issues by applying a tensegrity model to mammalian cells such as CHO cells. By applying a tensegrity force to such cells, the level of recombinant protein produced by the cells can be modulated. Summary of the invention [0005] The present invent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/67
CPCC12M35/06C12N15/67
Inventor R.皮帕里亚I.科雷亚
Owner ABBVIE INC
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