Multicomponent protein adsorption in supported cationic polyacrylamide hydrogels

Russell, Shawn M.; Carta, Giorgio

doi:10.1002/aic.10482

Cited by 32 publications

(27 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The introduction of labels reduces the net charge on lysozyme, and this can change chromatographic behavior in gradient elution relative to that of the unlabeled protein (Russell et al, 2003). These effects appear not to be present in uptake experiments at the lower salt concentrations (100 mM and below) used here.…”

Section: Protein Adsorbent and Chemicalscontrasting

confidence: 45%

Effects of ionic strength on lysozyme uptake rates in cation exchangers. I: Uptake in SP Sepharose FF

Dziennik

Belcher

Barker

et al. 2005

Biotech & Bioengineering

View full text Add to dashboard Cite

Fluorescence scanning confocal microscopy was used in parallel with batch uptake and breakthrough measurements of transport rates to study the effect of ionic strength on the uptake of lysozyme into SP Sepharose FF. In all cases the adsorption isotherms were near-rectangular. As described previously, the intraparticle profiles changed from slow-moving self-sharpening fronts at low salt concentration, to fast-moving diffuse profiles at high salt concentration, and batch uptake rates correspondingly increased with increasing salt concentration. Shrinking core and homogeneous diffusion frameworks were used successfully to obtain effective diffusivities for the low salt and high salt conditions, respectively. The prediction of column breakthrough was generally good using these frameworks, except for low-salt uptake results. In those cases, the compressibility of the stationary phase coupled with the shrinking core behavior appears to reduce the mass transfer rates at particle-particle contacts, leading to shallower breakthrough curves. In contrast, the fast uptake rates at high ionic strength appear to reduce the importance of mass transfer limitations at the particle contacts, but the confocal results do show a flow rate dependence on the uptake profiles, suggesting that external mass transfer becomes more limiting at high ionic strength. These results show that the complexity of behavior observable at the microscopic scale is directly manifested at the column scale and provides a phenomenological basis to interpret and predict column breakthrough. In addition, the results provide heuristics for the optimization of chromatographic conditions.

show abstract

Section: Protein Adsorbent and Chemicalscontrasting

confidence: 45%

Effects of ionic strength on lysozyme uptake rates in cation exchangers. I: Uptake in SP Sepharose FF

Dziennik

Belcher

Barker

et al. 2005

Biotech & Bioengineering

View full text Add to dashboard Cite

show abstract

“…Russell and Carta [57] and Teske et al [58] have recently shown that a chromatographic separation of labeled and unlabeled protein molecules is in fact readily obtained in gradient and isocratic ion exchange chromatography. While an exact comparison of the chromatographic selectivity with that used in the simulations is not possible because of the vastly different conditions, these experimental observations and the striking agreement of CLSM and predicted results suggest that the concentration overshoots observed in single protein CLSM uptake experiments are actually artifacts.…”

Section: Confocal Microscopymentioning

confidence: 99%

Protein Mass Transfer Kinetics in Ion Exchange Media: Measurements and Interpretations

2005

View full text Add to dashboard Cite

The paper gives a summary of successful methods for measuring protein mass transfer kinetics in ion exchange matrices along with models needed to interpret experimental results. Both macroscopic methods (isocratic elution, gradient elution, batch adsorption, frontal analysis) and microscopic methods are considered. In all cases the main focus is the determination of the effective intraparticle diffusivity in order to permit a comparison of different stationary phases and provide a basis for predicting chromatographic process performance. Experimental results for representative systems are evaluated alongside the experimental and modeling aspects. Practical criteria for the selection of experimental conditions and the application of different models are discussed along with the advantages and disadvantages of the various approaches.

show abstract

“…Results from direct confocal microscopy measurement corresponded well to the data obtained from indirect fluidphase measurements. However, other authors reported that the techniques used in CLSM, particularly the need to use dye-labelled protein to achieve protein visualisation, can alter the adsorption properties of the protein leading to artefacts [8,9]. In addition, it was shown that competition between dye-labelled and native protein molecules for the adsorption sites results in apparent adsorption patterns that exhibit either overshoots when the native protein is more favourably adsorbed, or smooth profiles when adsorption of the labelled protein is favoured [10 -13].…”

Section: Introductionmentioning

confidence: 97%

Confocal microscopy study of uptake kinetics of α‐lactalbumin and β‐lactoglobulin onto the cation‐exchanger SP Sepharose FF

El‐Sayed¹,

Chase²

2009

J of Separation Science

View full text Add to dashboard Cite

Confocal laser scanning microscopy (CLSM) was used to study single- and two-component protein uptake for alpha-lactalbumin (ALA) and beta-lactoglobulin (BLG), as models for whey proteins, to SP Sepharose FF at pH 3.7 during batch experiments in a finite bath. By coupling a fluorescent dye with the protein molecule, the penetration into individual adsorbent particles at different times during batch uptake was visualised. In a single-component system, BLG penetrated fast into the adsorbent beads and gradually filled them in a shell-wise fashion, while adsorption of ALA was mostly confined to the outer shells of the adsorbent. For the two-component studies, the results showed that ALA was able to displace BLG despite its lower affinity to the adsorbent under the employed conditions. CLSM results were then compared both qualitatively and quantitatively to their counterparts obtained in traditional experiments by indirect measurements of the protein concentration in the fluid phase. A novel quantitative approach was undertaken by modifying the simple kinetic rate model traditionally used to determine the kinetic rate constant, k(1), for batch uptake experiments, in order to describe batch uptake kinetics based on CLSM data. Although BLG results were in good agreement, there was a discrepancy in ALA results.

show abstract

Multicomponent protein adsorption in supported cationic polyacrylamide hydrogels

Cited by 32 publications

References 27 publications

Effects of ionic strength on lysozyme uptake rates in cation exchangers. I: Uptake in SP Sepharose FF

Effects of ionic strength on lysozyme uptake rates in cation exchangers. I: Uptake in SP Sepharose FF

Protein Mass Transfer Kinetics in Ion Exchange Media: Measurements and Interpretations

Confocal microscopy study of uptake kinetics of α‐lactalbumin and β‐lactoglobulin onto the cation‐exchanger SP Sepharose FF

Contact Info

Product

Resources

About