Ultra scaledown to predict filtering centrifugation of secreted antibody fragments from fungal broth

Boulding, Natalie; Yim, Sung-Sam; Keshavarz-Moore, E; Shamlou, P. Ayazi; Berry, Mark J.

doi:10.1002/bit.10306

Cited by 12 publications

(10 citation statements)

References 15 publications

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“…It is significant that a non‐mechanical extraction method may be applicable at large scale for extraction of the secreted form of IgG, as this may offer economic savings at this initial stage of downstream processing. Furthermore, the application of preliminary ultra‐scale‐down techniques for large‐scale processing optimization is essential in order to save time and costs (Boulding et al ., 2002; Lee et al ., 2002; Neal et al ., 2003).…”

Section: Discussionmentioning

confidence: 99%

Considerations for extraction of monoclonal antibodies targeted to different subcellular compartments in transgenic tobacco plants

Hassan

Dolleweerd

Ioakeimidis

et al. 2008

Plant Biotechnology Journal

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SummaryMonoclonal antibody production from transgenic tobacco plants offers many advantages over other heterologous production systems, creating the prospect of production at a scale that will allow new prophylactic and therapeutic applications in global human and animal health. However, information on the major processing factors to consider for large-scale purification of antibodies from transgenic plants is currently limited, and is in urgent need of attention. The purpose of this project was to investigate methods for the initial extraction of recombinant immunoglobulin G (IgG) antibodies from transgenic tobacco leaf tissue.Three different transgenic plant lines were studied in order to establish the parameters for optimal extraction of monoclonal antibodies that accumulate in the apoplasm, at the plasma membrane or within the endoplasmic reticulum. For each transgenic line, seven techniques for physical extraction were compared. The factors that determine the optimal extraction of antibodies from plants have a direct influence on the initial choice of expression strategy, and so must be considered at an early stage. The use of small-scale techniques that are applicable to large-scale purification was a particularly important consideration. The optimal extraction technique varied with the target location of IgG in the plant cell, and the dependence of antibody yield on the physical extraction methodology employed, the pH of the extraction buffer and the extraction temperature was demonstrated in each case. The addition of detergent to the extraction buffer may improve the yield, but this was found to be dependent on the site of accumulation of IgG within the plant cell.

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Section: Discussionmentioning

confidence: 99%

Considerations for extraction of monoclonal antibodies targeted to different subcellular compartments in transgenic tobacco plants

Hassan

Dolleweerd

Ioakeimidis

et al. 2008

Plant Biotechnology Journal

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“…In addition, experimental data were gathered to validate simulations made by CFX [9]. The package is now routinely in our studies on the assessment of flow in complex process geometries [10,11,12] including the feed zones described in this paper for the disc-stack centrifuge [6] and the multichamber-bowl centrifuge [1].…”

Section: Theoretical Considerationsmentioning

confidence: 99%

“…Maximum energy dissipation rates calculated using CFD N.D. not determined. a Based on data presented elsewhere for similar yeast protein precipitates[10] assuming the same level of shear damage observed.…”

mentioning

confidence: 99%

Performance prediction of industrial centrifuges using scale-down models

Boychyn

Yim

Bulmer

et al. 2003

Bioprocess Biosyst Eng

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122

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Computational fluid dynamics was used to model the high flow forces found in the feed zone of a multichamber-bowl centrifuge and reproduce these in a small, high-speed rotating disc device. Linking the device to scale-down centrifugation, permitted good estimation of the performance of various continuous-flow centrifuges (disc stack, multichamber bowl, CARR Powerfuge) for shear-sensitive protein precipitates. Critically, the ultra scale-down centrifugation process proved to be a much more accurate predictor of production multichamber-bowl performance than was the pilot centrifuge.

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“…Microfiltration is an attractive separation option, particularly for protein precipitates in suspension, which may show susceptibility to shear‐induced damage in centrifuges [20,21]. It is noted that shear rates at the surface of the microfiltration membrane can be high.…”

Section: Resultsmentioning

confidence: 99%

“…There are two possible reasons for this. The first is the potential shear‐associated damage to the product antibody in the high‐shear zones in an industrial centrifuge [1,20,21]. The second is the capacity of the continuous microfiltration to increase the overall process yield by integrating several operations (concentration, diafiltration, washing and buffer exchange) in a single piece of equipment.…”

Section: Resultsmentioning

confidence: 99%

Separation of immunoglobulin G precipitate from contaminating proteins using microfiltration

Neal¹,

Francis²,

Shamlou³

et al. 2004

Biotech and App Biochem

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A small-scale stirred-cell device was used to separate an antivenom antibody precipitate from a suspension containing contaminating soluble proteins. The device has a total volume of 200 ml and is equipped with a filter membrane with a cut-off size of 3 microm. About 90% of the antibody-precipitate particles in the feed were 29 microm or smaller, and the concentration of solids was 12% (w/w). The microfiltration cell was operated in a constant-volume (continuous/diafiltration) mode, and its performance was compared with an industrial disc-stack centrifuge currently used in the manufacture of antibody precipitate. In terms of product purity, the separation performance of the microfiltration operation was found to be comparable with the disc-stack centrifuge, whereas the overall yield was 10% better than that obtained from the centrifuge. This was attributed to the ability of microfiltration to reduce material losses by integrating a number of operations in a single piece of equipment. These included separation, concentration and buffer exchange, as well as dissolution and final recovery of the antibody in an appropriate buffer. The results obtained from the stirred cell are potentially scaleable, and dynamic microfiltration is shown to be an attractive process option.

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Ultra scaledown to predict filtering centrifugation of secreted antibody fragments from fungal broth

Cited by 12 publications

References 15 publications

Considerations for extraction of monoclonal antibodies targeted to different subcellular compartments in transgenic tobacco plants

Considerations for extraction of monoclonal antibodies targeted to different subcellular compartments in transgenic tobacco plants

Performance prediction of industrial centrifuges using scale-down models

Separation of immunoglobulin G precipitate from contaminating proteins using microfiltration

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