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Perfusion Cell Culture

Perfusion, or upstream continuous bioprocessing, has been practiced in cell culture since the 1980s but with limited adoption. Increasing use of innovative technologies, turnkey systems, cell culture supplements and single-use assemblies has changed the paradigm, making perfusion cell culture a viable platform for continuous processing. 

long-term perfusion

Perfusion is upstream processing which retains cells inside the bioreactor while continually removing cell waste products and media depleted of nutrients by cell metabolism. Fresh media is provided to the cells at the same rate as the spent media is removed. The most common means to achieve perfusion is the use of hollow fiber filtration.

Background

Perfusion has been practiced in cell culture since the 1980s but with limited adoption due to complex or unreliable equipment and scale-up challenges associated with cell retention.  Separate advances in cell line engineering, media composition, and bioreactor design generated magnitude-fold increases in titers for batch and fed-batch modes, thereby reducing the need for adoption of perfusion technology during that time. 

Recently, however, the need for alternative manufacturing strategies that can boost efficiency and productivity while reducing costs has led to renewed interest in perfusion technology.

The introduction of the XCell™ ATF System, using novel alternating tangetial flow filtration,  has changed the paradigm, making perfusion cell culture a viable platform for modern continuous processing.

As the provider of the broadest range of filtration technologies for bioprocessing, Repligen also offers KrosFlo® systems using traditional tangential flow filtration,  as well as potent cell culture supplements and custom flow path assemblies.

Brief History of Perfusion Biomanufacturing

Advances in long-term perfusion

Alternating Tangential Flow (ATF) Filtration

An award-winning technology, ATF allows the removal of spent media while keeping cells in culture.  Applied using an XCell™ ATF device attached to a bioreactor, ATF minimizes cell shear and keeps cells in constant equilibrium with bioreactor contents. This results in faster cell growth at higher densities with higher productivity.  Today, the XCell™ ATF device, in single-use or reusable format, is the leading perfusion device for mAb and rProtein production.

Tangential Flow Filtration (TFF) Systems

Turnkey, easy-to-use, cost-effective and scalable hollow fiber systems for tangential flow filtration (TFF), single-use filter modules and state-of-the-art hollow fiber membranes enable process and cost efficiencies in upstream perfusion. KrosFlo® KML 100 and KPS TFF Systems are fully integrated and ready to use out of the box, the systems include precise and reproducible control and documentation of all TFF processes.

Single-use custom flow paths 

Ready-to-use connectivity between any bioreactor and the cell retention device facilitates the adoption of perfusion technologies.  Defined through an engineering design process, Pro-Connex® Flow Paths are customized, configurable and complete assemblies for tangential flow filtration. The assemblies are sterile and manufactured in compliance with ISO guidelines in an ISO 7 clean room.

Pro-Connex® Flow Paths

Potent cell culture supplements

Repligen developed and manufactures cell culture supplements that provide the benefits of serum supplementation while maintaining an animal-free process.

Cell culture supplements

Industry Best Practices

As a technology leader in bioprocessing, Repligen has compiled articles and essays from thought leaders on topics pertaining to continuous bioprocessing into easy-to-access e-books. These e-books summarize current thoughts on this paradigm shift in bioprocessing , the technical challenges, existing solutions and the promised opportunities.

The opinions expressed in these books are those of the authors and not of Repligen Corporation.

Continuous Bioprocessing: Industry Best Practices

E-book Volume 2
February 2018

Compiled by John Bonham-Carter, Repligen

Chapters

  1. Introduction to Continuous Bioprocessing: Current Status and Trends
    Eric S. Langer, BioPlan Associates, Inc. and and Ronald A. Rader
  2. How to Develop a Perfusion Process
    Véronique Chotteau, Royal Institute of Technology (KTH)
  3. Strategies for development of perfusion media
    Delia Lyons, MilliporeSigma and Jochen Sieck, Merck KGaA
  4. Perfusion Application in Mobius® Single-use Bioreactors
    Michael Cunningham and Douglas Rank, MIlliporeSigma
  5. XCell™ Perfusion Technology Delivers High Throughput: Efficient Seed Train Process with Improved Fed-Batch Production Output
    W. Roy Lin, Shashi Kudugunti, Mario Sinani, Tim Erlandson and Dr. James Rusche, Repligen
  6. Product Quality Considerations of Perfusion Processes
    Thomas Villiger, Daniel Karst and Massimo Morbidelli, ETH Zurich
  7. Impurity Monitoring as Novel PAT Tool for Continuous Biopharmaceutical Processes
    Vignesh Rajamanickam, Dr. Patrick Sagmeister, Oliver Spadiut, and Christoph Herwig, TU Wien Austria
  8. Insect Cells and ATF
    Willem Adriaan (Wian) de Jongh, PhD, ExpreS2ion Biotechnologies
  9. Scale up and Technology Transfer in a Continuous Processing Environment
    Sadettin S. Ozturk, Ph.D., Massbiologics
  10. Continuous Multicolumn Chromatography Processes
    Marc Bisschops, Pall Corporation
  11. Impact of Single‑use Technology on Continuous Bioprocessing
    William G. Whitford, GE Healthcare
  12. Continuous Bioprocessing's Impact on Facility Design
    Morten Munk, NNE A/S

 

     Continuous Processing E-Book Volume 2

Continuous Bioprocessing: Current Practice and Future Potential

E-book Volume 1
2014

Compiled by John Bonham-Carter, Repligen

Chapters

  1. Introduction to Continuous Manufacturing:Technology Landscape and Trends
    Eric S. Langer and Ronald A. Rader, BioPlan Associates
  2. A Brief History of Perfusion Biomanufacturing
    Jerry Shevitz and John Bonham-Carter, Refine Technology
  3. Bioreactor Configuration and Operation
    Christel Fenge, Jörg Weyand, Gerhard Greller, Thorsten Adams, Sartorius Stedim Biotech
  4. How to Develop a Perfusion Process
    Véronique Chotteau, Royal Technical University, KTH
  5. Case Study: Optimized Perfusion
    Shaun Eckerle, Gallus BioPharmaceuticals
  6. Process Intensification Approaches for Cost Sensitive Protein Applications
    Willem Adriaan de Jongh, Expres2ion Biotechnologies
  7. Impact of Single-use Technology on Continuous Biorocessing
    William G. Whitford, Thermo Fisher Scientific
  8. Continuous Multicolumn Chromatography Processes
    Marc Bisschops, Tarpon Biosystems
  9. Continuous Processes: Economic Evaluation
    Andrew Sinclair and Andrew Brown, BioPharm Services
  10. Vision: Integrating Upstream and Downstream in a Fully Continuous Facility
    Tim Johnson, Genzyme

 

     Continuous processing e-book Volume 1