A Continuous Cell Centrifuge for Lab Scale Perfusion Processes of Mammalian Cells

Chatzisavido, Nathalie; Björling, Torsten; Fenge, Christel; Boork, Sarah; Lindner-Olsson, Elisabeth; Apelman, Stig

doi:10.1007/978-94-011-0848-5_70

Cited by 10 publications

(6 citation statements)

References 2 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As already shown, the dead cells in the supernatant came from the bioreactor. Evidence of a selective dead‐cell bleed‐out from a bioreactor using centrifugation has been previously reported (Apelman, 1991; Apelman and Björling, 1991; Björling and Malström, 1991; Chatzisavido et al, 1993; Wie et al, 1991). In those studies, dead cells were shown to settle somewhat slower than viable cells, and dead‐cell removal was noted to be necessary to reduce lysis‐triggered release of proteolytic enzymes inhibiting viable‐cell growth and productivity.…”

Section: Resultsmentioning

confidence: 77%

“…This paper focuses on the evaluation of a bench‐top centrifugal separator, the first prototype of the Centritech Lab (Centritech AB, Norsborg, Sweden). Because few results have been reported on the use of the Centritech Lab (Chatzisavido et al, 1993; Tokashiki and Takamatsu, 1993), both its technical performance and the effect of its use on culture behavior were evaluated. Operating parameters, component durability, separation efficiency, and monoclonal‐antibody recovery were characterized.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Use of the Centritech Lab Centrifuge for Perfusion Culture of Hybridoma Cells in Protein‐Free Medium

Johnson¹,

Lanthier²,

Massie³

et al. 1996

Biotechnology Progress

View full text Add to dashboard Cite

As part of an effort to develop a suspension-culture perfusion-based process with high flow rate without the fouling and antibody retention inherent to filter-based cell-separation devices, we have evaluated and contributed to the development of the Centritech Lab centrifuge for the perfusion culture of hybridoma cells in protein-free medium. Culture start-ups showed that cell growth and monoclonal-antibody (MAb) production rates were similar in both a spinner flask and continuous centrifugation coupled to a bioreactor. The centrifuge efficiently separated viable cells from dead ones. Viable-cell recoveries were never below 98%, whereas dead-cell recoveries were usually around 80%. The cell content of the centrifuge supernatant and concentrate was strongly determined by the total amount of cells, viable and dead, in the culture broth, but an influence of the centrifugation parameters (feed rate, times of separation and discharge, and rotor speed) was observed. This understanding of the separation process inside the centrifuge is important and may apply to other similar devices. Monoclonal antibodies were not retained in the bioreactor during centrifugation perfusion. However, whereas similar growth rates were obtained in perfusion cultures using either continuous centrifugation or filtration, MAb concentrations were 35% lower in the former case. Utilization of the centrifuge in an intermittent fashion decreased the daily cell residence time outside the bioreactor, the daily pelleted-cell residence time in the centrifuge, and the frequency of cell passage to the centrifuge. This led to higher viable-cell numbers in the bioreactor and an accompanying increase in MAb concentrations, 225-250 mg of IgM L-1, equal to the performance of filter-based perfusion systems with the same cell line. It was hypothesized that having cells periodically packed at the bottom of the centrifuge insert (up to 800 x 10(6) cells mL-1) is deleterious to the culture by exposing the pelleted cells to prolonged nutrient limitations.

show abstract

Section: Resultsmentioning

confidence: 77%

Section: Introductionmentioning

confidence: 99%

Use of the Centritech Lab Centrifuge for Perfusion Culture of Hybridoma Cells in Protein‐Free Medium

Johnson¹,

Lanthier²,

Massie³

et al. 1996

Biotechnology Progress

View full text Add to dashboard Cite

show abstract

“…Described in the early 1990s (Apelman 1992;Apelman and Bjorling 1991;Chatzisavido et al 1993), it has been adopted in some commercial perfusion processes for the production of labile proteins, e.g. Shire (two CELL Centritech systems connected to a 2,000 L bioreactor), Swedish Orphan Biovitrum (D-domain deleted factor VIII Refacto® manufacturing for Pfizer at 500 L scale).…”

Section: Centrifugementioning

confidence: 99%

Perfusion Processes

Chotteau

2014

Cell Engineering

View full text Add to dashboard Cite

The interest for perfusion is increasing nowadays. This new focus has emerged from a synergy of a demand for disposable equipment and the availability of robust cell separation device, as well as the need for higher flexibility and lower investment cost. The cell separation devices mostly used today are based on filtration, i.e. alternating flow filtration, tangential flow filtration, spin-filter, or acceleration/gravity, i.e. inclined settler, centrifuge, acoustic settler. This paper gives an introduction to the basic concepts of perfusion and its practical implementation. It reviews the actual cell separation devices and describes the approaches used in the field to develop and optimize the perfusion processes.

show abstract

“…It can also be noticed that separation inserts have been approved by the FDA. Several investigators used these centrifuges to grow CHO (Apelman, 1992;Arai et al, 1993;Chatzisavido et al, 1994) and SP2/0 cells (Johnson et al, 1996) in perfusion. Results of cell cultures are given in Table II.…”

Section: Centrifugesmentioning

confidence: 99%

Potential of cell retention techniques for large‐scale high‐density perfusion culture of suspended mammalian cells

Voisard

Meuwly

Ruffieux

et al. 2003

Biotech & Bioengineering

244

150

View full text Add to dashboard Cite

This review focuses on cultivation of mammalian cells in a suspended perfusion mode. The major technological limitation in the scaling-up of these systems is the need for robust retention devices to enable perfusion of medium as needed. For this, cell retention techniques available to date are presented, namely, cross-flow filters, hollow fibers, controlled-shear filters, vortex-flow filters, spin-filters, gravity settlers, centrifuges, acoustic settlers, and hydrocyclones. These retention techniques are compared and evaluated for their respective advantages and potential for large-scale utilization in the context of industrial manufacturing processes. This analysis shows certain techniques have a limited range of perfusion rate where they can be implemented (most microfiltration techniques). On the other hand, techniques were identified that have shown high perfusion capacity (centrifuges and spin-filters), or have a good potential for scale-up (acoustic settlers and inclined settlers). The literature clearly shows that reasonable solutions exist to develop large-scale perfusion processes.

show abstract

A Continuous Cell Centrifuge for Lab Scale Perfusion Processes of Mammalian Cells

Cited by 10 publications

References 2 publications

Use of the Centritech Lab Centrifuge for Perfusion Culture of Hybridoma Cells in Protein‐Free Medium

Use of the Centritech Lab Centrifuge for Perfusion Culture of Hybridoma Cells in Protein‐Free Medium

Perfusion Processes

Potential of cell retention techniques for large‐scale high‐density perfusion culture of suspended mammalian cells

Contact Info

Product

Resources

About