Direct Quantification of Intraparticle Protein Diffusion in Chromatographic Media

Schröder, Magnus; Lieres, Eric von; Hubbuch, Jürgen

doi:10.1021/jp0542726

Cited by 34 publications

(21 citation statements)

References 31 publications

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“…If that can be done at the microfluidic level it would ease this. Also at very small scale, studies of intraparticle protein diffusion and uptake in micro-columns using confocal laser scanning microscopy have allowed data to be collected at a very small scale which agree with values obtained at large scales (Dziennik et al, 2005;Schroder et al, 2006). Siu et al (2007) showed that this approach could be used to assess fouling and that the miniature column format, bed volume 66 mL with flow through, was better for prediction than a batch test.…”

Section: Microwell-based Studiessupporting

confidence: 58%

Micro biochemical engineering to accelerate the design of industrial‐scale downstream processes for biopharmaceutical proteins

Titchener‐Hooker

Dunnill

Hoare

2008

Biotech & Bioengineering

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The article examines how a small set of easily implemented micro biochemical engineering procedures combined with regime analysis and bioprocess models can be used to predict industrial scale performance of biopharmaceutical protein downstream processing. This approach has been worked on in many of our studies of individual operations over the last 10 years and allows preliminary evaluation to be conducted much earlier in the development pathway because of lower costs. It then permits the later large scale trials to be more highly focused. This means that the risk of delays during bioprocess development and of product launch are reduced. Here we draw the outcomes of this research together and illustrate its use in a set of typical operations; cell rupture, centrifugation, filtration, precipitation, expanded bed adsorption, chromatography and for common sources, E. coli, two yeasts and mammalian cells (GS-NSO). The general approach to establishing this method for other operations is summarized and new developments outlined. The technique is placed against the background of the scale-down methods that preceded it and complementary ones that are being examined in parallel. The article concludes with a discussion of the advantages and limitations of the micro biochemical engineering approach versus other methods.

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Section: Microwell-based Studiessupporting

confidence: 58%

Micro biochemical engineering to accelerate the design of industrial‐scale downstream processes for biopharmaceutical proteins

Titchener‐Hooker

Dunnill

Hoare

2008

Biotech & Bioengineering

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“…Finally, the diffusion coefficients used in this study were taken from literature (Dziennik et al, 2003;Coffman et al, 1997;Cussler, 1997). In a recent study Schröder et al (2006) determined the diffusion coefficient for lysozyme labeled with Cy5 in Sepharose 6 FF to 6.07 Â 10 À7 cm 2 /s which is slightly higher than assumed in this study. The influence of this should, however, be negligible within the scope of his study.…”

Section: Sma Parameter Of Bio-conjugatesmentioning

confidence: 49%

“…In the calculation, we assumed that the external concentration of labeled and native protein was an ideal step function as the experiment begins. The experimental studies, however, were conducted in a packed bed and the protein concentration at the surface of the observed particle is not a perfect step (Schröder et al, 2006) and the labeled and native concentration changes may not occur simultaneously due to the separation occurring in the bed. The incorrect prediction that the overshoot magnitude at 6 mM ionic strength is greater than at 50 mM ionic strength are at least partly due to neglecting the kinetics of adsorption and desorption of the protein.…”

Section: Model Prediction Versus Clsm Datamentioning

confidence: 99%

Competitive adsorption of labeled and native protein in confocal laser scanning microscopy

Teske

Lieres

Schröder

et al. 2006

Biotech & Bioengineering

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Confocal laser scanning microscopy has been previously applied to the study of protein uptake in porous chromatography resins. This method requires labeling the protein with a fluorescent probe. The labeled protein is then diluted with a large quantity of native protein so that the fluorescence intensity is a linear function of the labeled protein concentration. Ideally, the attachment of a fluorescent probe should not affect the affinity of the protein for the stationary phase; however, recent experimental work has shown that this assumption is difficult to satisfy. In the present study, we present a mathematical model of protein diffusion and adsorption in a single adsorbent particle. The differences in adsorption behavior of labeled and native protein are accounted for by treating the system as a two-component system (labeled and native protein) described by the steric mass action isotherm (SMA). SMA parameters are regressed from experimental linear gradient elution data for lysozyme and lysozyme-dye conjugates (for the fluorescent dyes Cy3, Cy5, Bodipy FL, and Atto635). When the regressed parameters are employed in the model, an overshoot in the labeled lysozyme concentration is predicted for Cy5- and Bodipy-labeled lysozyme, but not for Atto635-labeled lysozyme. The model predictions agree qualitatively well with recent work showing the dependence of the concentration overshoot on the identity of the attached dye and provide further evidence that the overshoot is likely caused by the change of binding characteristics due to the fluorescent label.

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“…Immobilization is the net result of various conformational changes that occur within and outside the enzyme surface. Immobilization could cause altered enzyme activity (Schröder et al, 2006). The formation of individual biomolecules and their orientation on surfaces can easily be monitored by different microscopic and spectroscopic techniques.…”

Section: Recent Tools and Technique For Structural Determination Of Imentioning

confidence: 99%

How enzymes are adsorbed on soil solid phase and factors limiting its activity: A Review

Datta¹,

Anand²,

Moulick³

et al. 2017

International Agrophysics

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A b s t r a c t. A majority of biochemical reactions are often catalysed by different types of enzymes. Adsorption of the enzyme is an imperative phenomenon, which protects it from physical or chemical degradation resulting in enzyme reserve in soil. This article summarizes some of the key results from previous studies and provides information about how enzymes are adsorbed on the surface of the soil solid phase and how different factors affect enzymatic activity in soil. Many studies have been done separately on the soil enzymatic activity and adsorption of enzymes on solid surfaces. However, only a few studies discuss enzyme adsorption on soil perspective; hence, we attempted to facilitate the process of enzyme adsorption specifically on soil surfaces. This review is remarkably unmatched, as we have thoroughly reviewed the relevant publications related to protein adsorption and enzymatic activity. Also, the article focuses on two important aspects, adsorption of enzymes and factors limiting the activity of adsorbed enzyme, together in one paper. The first part of this review comprehensively lays emphasis on different interactions between enzymes and the soil solid phase and the kinetics of enzyme adsorption. In the second part, we encircle various factors affecting the enzymatic activity of the adsorbed enzyme in soil.

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Direct Quantification of Intraparticle Protein Diffusion in Chromatographic Media

Cited by 34 publications

References 31 publications

Micro biochemical engineering to accelerate the design of industrial‐scale downstream processes for biopharmaceutical proteins

Micro biochemical engineering to accelerate the design of industrial‐scale downstream processes for biopharmaceutical proteins

Competitive adsorption of labeled and native protein in confocal laser scanning microscopy

How enzymes are adsorbed on soil solid phase and factors limiting its activity: A Review

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