Separation of Whey Protein Aggregates by Means of Continuous Centrifugation

Haller, Nicole; Kulozik, Ulrich

doi:10.1007/s11947-019-02275-1

Cited by 20 publications

(7 citation statements)

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“…Although centrifugation is the most attractive option for washing of precipitates at small scale, the dense packing of the precipitate / flocs can create challenges in both product recovery and washing in large‐scale biomanufacturing 9 . For example, Haller and Kulozik 13 observed significant particle breakdown during continuous centrifugation of α‐lactalbumin, with the adhesiveness of the concentrated precipitate leading to high yield loss during discharge from the centrifuge. These problems can be largely eliminated using MF.…”

Section: Introductionmentioning

confidence: 99%

Enhanced filtration performance usingfeed‐and‐bleedconfiguration for purification of antibody precipitates

Zhao

Chen

Andini

et al. 2020

Biotechnology Progress

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Precipitation can be used for the initial purification of monoclonal antibodies (mAbs), with the soluble host cell proteins removed in the permeate by tangential flow microfiltration. The objective of this study was to examine the use of a feed-andbleed configuration to increase the effective conversion (ratio of permeate to feed flow rates) in the hollow fiber module to enable more effective washing of the precipitate. Experiments were performed using human serum Immunoglobulin G (IgG) precipitates formed with 10 mM zinc chloride and 7 wt% polyethylene glycol. The critical flux was evaluated as a function of the shear rate and IgG concentration, with the resulting correlation used to predict conditions that can achieve 90% conversion in a single pass with minimal fouling. Experimental data for both the start-up and steady-state performance are in good agreement with model calculations. These results were used to analyze the performance of an enhanced continuous precipitation-microfiltration process using the feed-and-bleed configuration for the initial capture / purification of a mAb product.

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Section: Introductionmentioning

confidence: 99%

Enhanced filtration performance usingfeed‐and‐bleedconfiguration for purification of antibody precipitates

Zhao

Chen

Andini

et al. 2020

Biotechnology Progress

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“…Therefore, the βLG redissolved in distilled water had a pH of 4.77 due to the missing buffering capacity of the aqueous phase. The method is described in [22]. The βLG was highly pure (<0.05% lactose, minerals <0.13%), the moisture content was <4%, the degree of nativity was >99%, with 96.8% protein on dry matter, the content of βLG in the dry matter was >99%.…”

Section: Methodsmentioning

confidence: 99%

Influence of Thermomechanical Treatment and Ratio of β-Lactoglobulin and α-Lactalbumin on the Denaturation and Aggregation of Highly Concentrated Whey Protein Systems

Quevedo

Kulozik

Karbstein

et al. 2020

Foods

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The influence of thermomechanical treatment (temperature 60 °C–100 °C and shear rate 0.06 s−1–50 s−1) and mixing ratio of β-lactoglobulin (βLG) and α-lactalbumin (αLA) (5:2 and 1:1) on the denaturation and aggregation of whey protein model systems with a protein concentration of 60% and 70% (w/w) was investigated. An aggregation onset temperature was determined at approx. 80 °C for both systems (5:2 and 1:1 mixing ratio) with a protein concentration of 70% at a shear rate of 0.06 s−1. Increasing the shear rate up to 50 s−1 led to a decrease in the aggregation onset temperature independent of the mixing ratio. By decreasing the protein concentration to 60% in unsheared systems, the aggregation onset temperature decreased compared to that at a protein concentration of 70%. Furthermore, two significantly different onset temperatures were determined when the shear rate was increased to 25 s−1 and 50 s−1, which might result from a shear-induced phase separation. Application of combined thermal and mechanical treatment resulted in overall higher degrees of denaturation independent of the mixing ratio and protein concentration. At the conditions applied, the aggregation of the βLG and αLA mixtures was mainly due to the formation of non-covalent bonds. Although the proportion of disulfide bond aggregation increased with treatment temperature and shear rate, it was higher at a mixing ratio of 5:2 compared to that at 1:1.

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“…In order to obtain a measure for the deposit porosity, the equilibrium particle volume fraction in the sediment Φ base can be calculated based on initial particle volume fraction Φ 0 , initial sample height H 0 , radial position of the cuvette bottom R max and centrifugal acceleration a, according to Equation (5) [24,25].…”

Section: Compressibility Analysismentioning

confidence: 99%

Determination of Compressibility and Relaxation Behavior of Yeast Cell Sediments by Analytical Centrifugation and Comparison with Deposit Formation on Membrane Surfaces

Weinberger

Kulozik

2022

Membranes

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Separation of cells from produced biomolecules is a challenging task in many biotechnological downstream operations due to deposit formation of the retained cells, affecting permeation of the target product. Compression and relaxation behavior of cell deposits formed during filtration are important factors affecting operational performance. The determination of these factors by flux or pressure stepping experiments is time- and labor-intensive. In this work, we propose a screening method by analytical centrifugation, which is capable of detecting small differences in compression and relaxation behavior induced by milieu changes, using a model system comprised of washed and unwashed yeast cells in the presence or absence of bovine serum albumin as a model target protein. The main effects observed were firstly the impact of pH value, affecting interaction of bovine serum albumin and yeast cells especially close to the isoelectric point, and secondly the effect of washing the yeast cells prior to analysis, where the presence of extracellular polymeric substances led to higher compressibility of the deposited cells. By comparing and validating the obtained results with dead-end filtration trials, the stabilizing role of bovine serum albumin in deposits formed at low pH values due to interactions with the yeast cells was underlined.

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Separation of Whey Protein Aggregates by Means of Continuous Centrifugation

Cited by 20 publications

References 50 publications

Enhanced filtration performance usingfeed‐and‐bleedconfiguration for purification of antibody precipitates

Enhanced filtration performance usingfeed‐and‐bleedconfiguration for purification of antibody precipitates

Influence of Thermomechanical Treatment and Ratio of β-Lactoglobulin and α-Lactalbumin on the Denaturation and Aggregation of Highly Concentrated Whey Protein Systems

Determination of Compressibility and Relaxation Behavior of Yeast Cell Sediments by Analytical Centrifugation and Comparison with Deposit Formation on Membrane Surfaces

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