Crossflow microfiltration of recombinant Escherichia coli lysates after high pressure homogenization

Bailey, Stuart; Meagher, Michael M.

doi:10.1002/(sici)1097-0290(19971105)56:3<304::aid-bit8>3.3.co;2-y

Cited by 5 publications

(6 citation statements)

References 35 publications

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“…This relationship is reasonable and is supported qualitatively by Forman et al, who showed that a protein exhibited a sieving coefficient higher than 90% at a very low permeation flux of 3 Lmh (39). This is also corroborated by the work of Bailey and Meagher (40). The sieving coefficient through the membrane pores is evaluated by the traditional method based on solute partitioning coefficient, solvent transport parameters, and membrane characteristics as described in Zeman and Zydney (16).…”

Section: Theoretical Backgroundsupporting

confidence: 78%

Global Model for Optimizing Crossflow Microfiltration and Ultrafiltration Processes: A New Predictive and Design Tool

Baruah

Venkiteshwaran

Belfort

2008

Biotechnol Progress

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A global model and algorithm that predicts the performance of crossflow MF and UF process individually or in combination in the laminar flow regime is presented and successfully tested. The model accounts for solute polydispersity, ionic environment, electrostatics, membrane properties and operating conditions. Computer programs were written in Fortran 77 for different versions of the model algorithm that can optimize MF/UF processes rapidly in terms of yield, purity, selectivity, or processing time. The model is validated successfully with three test cases: separation of bovine serum albumin (BSA) from hemoglobin (Hb), capture of immunoglobulin (IgG) from transgenic goat milk by MF, and separation of BSA from IgG by UF. These comparisons demonstrate the capability of the global model to conduct realistic in silico simulations of MF and UF processes. This model and algorithm should prove to be an invaluable technique to rapidly design new or optimize existing MF and UF processes separately or in combination in both pressure-dependent and pressure-independent regimes.

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Section: Theoretical Backgroundsupporting

confidence: 78%

Global Model for Optimizing Crossflow Microfiltration and Ultrafiltration Processes: A New Predictive and Design Tool

Baruah

Venkiteshwaran

Belfort

2008

Biotechnol Progress

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“…Datar and Rosen (1987) even suggested a two-stage homogenizer or an auxillary equipment to reduce the viscosity of homogenates. However, repeated passage of cells through a homogenizer is undesirable due to the micronisation of the cell debris (Bailey and Meagher, 1997;Bailey et al, 1995;Keshavarz, 1991;Keshavarz et al, 1990) and its detrimental effect on clarification (Agerkvist and Enfors, 1990;Hutchinson et al, 2006), precipitation (Hagen et al, 1996), filtration (Lee et al, 2004;Shu-Sen, 1988;Tarleton and Wakeman, 1994), and chromatography (Grabski et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

Step change in the efficiency of centrifugation through cell engineering: co‐expression of Staphylococcal nuclease to reduce the viscosity of the bioprocess feedstock

Balasundaram

Nesbeth

Ward

et al. 2009

Biotech & Bioengineering

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Cell engineering to enable step change improvements in bioprocessing can be directed at targets other than increasing product titer. The physical properties of the process suspension such as viscosity, for example, have a major impact on various downstream processing unit operations. The release of chromosomal DNA during homogenization of Escherichia coli and its influence on viscosity is well-recognized. In this current article we demonstrate co-expression of Staphylococcus aureus nuclease in E. coli to reduce viscosity through auto-hydrolysis of nucleic acids. Viscosity reduction of up to 75% was achieved while the particle size distribution of cell debris was maintained approximately constant (d(50) = 0.5-0.6 microm). Critically, resultant step change improvements to the clarification performance under disc-stack centrifugation conditions are shown. The cell-engineered nuclease matched or exceeded the viscosity reduction performance seen with the addition of exogenous nuclease removing the expense and validation issues associated with such additions to a bioprocess. The resultant material dramatically altered performance in scale-down mimics of continuous disc-stack centrifugation. Laboratory scale data indicated that a fourfold reduction in the settling area of a disc-stack centrifuge can be expected due to a less viscous process stream achieved through nuclease co-expression with a recombinant protein.

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“…Figure shows the effect of homogenization on thawed cell broth. Homogenization at 500 bar for two passes is used as it reflects a typical process for complete cell disruption and total product release (Bailey and Maegher, ). As expected, there is a considerable shift towards smaller particle sizes with all aggregates above ∼10 µm (Fig.…”

Section: Resultsmentioning

confidence: 99%

Ultra scale‐down characterization of the impact of conditioning methods for harvested cell broths on clarification by continuous centrifugation—Recovery of domain antibodies from rec E. coli

Chatel¹,

Kumpalume

Hoare³

2013

Biotech & Bioengineering

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The processing of harvested E. coli cell broths is examined where the expressed protein product has been released into the extracellular space. Pre-treatment methods such as freeze–thaw, flocculation, and homogenization are studied. The resultant suspensions are characterized in terms of the particle size distribution, sensitivity to shear stress, rheology and solids volume fraction, and, using ultra scale-down methods, the predicted ability to clarify the material using industrial scale continuous flow centrifugation. A key finding was the potential of flocculation methods both to aid the recovery of the particles and to cause the selective precipitation of soluble contaminants. While the flocculated material is severely affected by process shear stress, the impact on the very fine end of the size distribution is relatively minor and hence the predicted performance was only diminished to a small extent, for example, from 99.9% to 99.7% clarification compared with 95% for autolysate and 65% for homogenate at equivalent centrifugation conditions. The lumped properties as represented by ultra scale-down centrifugation results were correlated with the basic properties affecting sedimentation including particle size distribution, suspension viscosity, and solids volume fraction. Grade efficiency relationships were used to allow for the particle and flow dynamics affecting capture in the centrifuge. The size distribution below a critical diameter dependant on the broth pre-treatment type was shown to be the main determining factor affecting the clarification achieved. Biotechnol. Bioeng. 2014;111: 913–924. © 2013 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

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Crossflow microfiltration of recombinant Escherichia coli lysates after high pressure homogenization

Cited by 5 publications

References 35 publications

Global Model for Optimizing Crossflow Microfiltration and Ultrafiltration Processes: A New Predictive and Design Tool

Global Model for Optimizing Crossflow Microfiltration and Ultrafiltration Processes: A New Predictive and Design Tool

Step change in the efficiency of centrifugation through cell engineering: co‐expression of Staphylococcal nuclease to reduce the viscosity of the bioprocess feedstock

Ultra scale‐down characterization of the impact of conditioning methods for harvested cell broths on clarification by continuous centrifugation—Recovery of domain antibodies from rec E. coli

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