Method for proliferation of cells within a bioreactor using a disposable pumphead and filter assembly

Inventors

Serway, David

Assignees

Repligen Corp

Abstract

A method for promoting biological activity uses a filter system to increase cell production of a fed batch bioreactor. The filter system cycles bioreactor fluid through a hollow fiber tangential flow filter which separates metabolic wastes (as well as proteins) from cells produced in bioreactor and returned to fed batch bioreactor, improving cell production in the fed batch bioreactor. The filter system includes a disposable pump and filter, and a reusable control system. The pump is a low shear gamma stable pump gently cycling bioreactor fluid through the filter with minimal damage to the cells produced in the bioreactor. The pumphead and hollow fiber tangential flow filter are disposable. The pump motor is part of the control system and is reusable. The pumphead and filter are provided as an assembled and pre-sterilized unit allowing simple and quick attachment to the fed batch bioreactor, and simple and quick disposal.

Core Innovation

The invention describes a fed-batch bioreactor proliferation method using a disposable, preassembled, presterilized tangential flow perfusion filter loop. The bioreactor fluid is recirculated through a hollow-fiber tangential flow filter to remove metabolic wastes, including associated proteins, while permeate is routed for disposal. The filtered retentate is returned to the bioreactor to improve cell density and protein productivity.

The system couples a disposable filtration and pump component to a reusable control system that recirculates fluid using a low-shear, gamma-stable, magnetically levitated disposable pumphead. The disposable pumphead and filter are provided as an assembled and presterilized unit and are configured for attachment to a vessel having feed tubing and return tubing. Aseptic connection/disconnection is enabled using sterile connectors, including self-closing needle-free sterile connectors.

Operation is supported with pressure/flow monitoring and non-invasive TMP pressure control, and includes priming and sterile connector functionality for aseptic operation. Defined filter architectures are included, including thin- or thick-wall hollow fiber tangential flow filters with depth filtration features, using mPES low protein-binding membranes and related chemistries. The document also describes elimination of autoclaving, and use of perfusion and clarification modes including seed and concentrated fed-batch.

Claims Coverage

The document provides two independent claims that define a tangential flow filtering system with two main inventive constructs: a disposable, preassembled, presterilized pumphead and tangential flow filter unit attachable to a vessel via aseptic connectors, and a reusable control system with a reusable pump motor coupled to the disposable pumphead. Additional claimed inventive features include permeate/waste-side tubing and, for one independent claim, a pressure sensor and a levitated rotor pumphead.

Across the independent claims, the core claim coverage is for a tangential flow filtering system that uses an assembled disposable, presterilized pumphead and tangential flow filter attachable to a bioreactor vessel feed and return lines via aseptic connectors, together with a reusable control system and coupleable reusable pump motor. One independent claim further requires a pressure sensor on the second fluid flow tubing and defines the disposable pumphead as having a levitated rotor, while also including waste/permeate-side tubing in fluid communication with the tangential flow filter.

Stated Advantages

Documented Applications