Determination of adsorption isotherm parameters for minor whey proteins by gradient elution preparative liquid chromatography

Faraji, Naeimeh; Zhang, Yan; Ray, Ajay K.

doi:10.1016/j.chroma.2015.08.004

Cited by 13 publications

(9 citation statements)

References 30 publications

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“…(8) to previously calibrated values in [26]. Broad statistical range in the determination of σ has been reported before in [15,16].…”

Section: Results Of the Mechanistic Model 421 Model Calibration Andmentioning

confidence: 98%

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Optimization of Lactoperoxidase and Lactoferrin Separation on an Ion-Exchange Chromatography Step

Faraji¹,

Zhang²,

Ray³

2017

Preprint

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Lactoperoxidase (LP), which is a high-value minor whey protein, has recently drawn extensive attention from research scientists and industry due to its multi-function and potential therapeutic applications. In this study, the separation and optimization of two similar-sized proteins, LP and lactoferrin (LF) were investigated using strong cation exchange column chromatography.Optimization was started with central composite design based experiments to characterize the importance of different decision variables. The three variables used in the optimization were flow rate, length of gradient and final salt concentration in the linear elution gradient step. The obtained empiric functional model represented the effect of the significant factors on the yield as the objective function. Afterwards, the calibrated mechanistic model was employed to predict accurate optimal set of variables. The optimal operating points were found and the results were compared with validation experiments. Predictions respecting yield confirmed a very good agreement with experimental results while keeping purity, a product quality characteristic, equal or above to a predefined value.

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“…(8) to previously calibrated values in [26]. Broad statistical range in the determination of σ has been reported before in [15,16].…”

Section: Results Of the Mechanistic Model 421 Model Calibration Andmentioning

confidence: 98%

“…In this section, the bed parameters of a 4.7 mL prepacked HiScreen TM SP Sepharose FF column characterized in [26] were set into the mechanistic model. The adsorption parameters with their confidence intervals are presented in Table 3.…”

Section: Results Of the Mechanistic Model 421 Model Calibration Andmentioning

confidence: 99%

Optimization of Lactoperoxidase and Lactoferrin Separation on an Ion-Exchange Chromatography Step

Faraji¹,

Zhang²,

Ray³

2017

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…The parameters were estimated based on DoE experiments in section 4.1 to keep the predictions over the similar design space. The steric factor (σ) had negligible influence on the fitting result; thus it was fixed during the optimization of Equation (8) to previously calibrated values in [32]. Broad statistical range in the determination of σ has been reported before in [21,22].…”

Section: Results Of Response Surface Modelingmentioning

confidence: 99%

“…In other words, quadratic RSM can only describe 66% of the variety in the experimental data. In this section, the bed parameters of a 4.7 mL prepacked HiScreen™ SP Sepharose FF column characterized in [32] were set into the mechanistic model. The adsorption parameters with their confidence intervals are presented in Table 3.…”

Section: Results Of Response Surface Modelingmentioning

confidence: 99%

Optimization of Lactoperoxidase and Lactoferrin Separation on an Ion-Exchange Chromatography Step

2017

Self Cite

View full text Add to dashboard Cite

Lactoperoxidase (LP), which is a high-value minor whey protein, has recently drawn extensive attention from research scientists and industry due to its multiplicity of function and potential therapeutic applications. In this study, the separation and optimization of two similar-sized proteins, LP and lactoferrin (LF) were investigated using strong cation exchange column chromatography. A two-step optimization strategy was developed for the separation of LP and LF. Optimization was started with central composite design-based experiments to characterize the influences of different decision variables, namely, flow rate, length of gradient, and final salt concentration in the linear elution gradient step on the yield of LP. This was followed by a more accurate optimization of ion-exchange chromatography (IEC) separation of LP and LF based on an experimentally verified chromatographic model. The optimal operating points were found and the results were compared with validation experiments. Predictions respecting yield confirmed a very good agreement with experimental results with improved product purity.

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“…Santos et al (2012) reported an anion exchange method for the isolation of BLG from whey concentrate. However, most of these reported chromatographic methods are either based on specialized separation media or multistep low purity processes or for the purification of only two proteins like LF and LPO (Faraji et al, 2015). Most importantly, reported whey protein isolation methods are of the multistep type, requiring the development of a single-step whey proteins isolation chromatography method for the detailed study of scale-up aspects.…”

Section: Introductionmentioning

confidence: 99%

Development and Optimization of a Single-Step Cation Chromatographic Whey Protein Fractionation Process: Evaluation and Comparison of Scale-Up Strategies

Ingle

Lali

2018

Braz. J. Chem. Eng.

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This manuscript presents evaluation of various strategies used for the scale-up of chromatographic purification systems. Towards that end, the manuscript provides development and optimization of a single-step whey protein concentrate (WPC) fractionation process at lab scale on a 1.0 × 19 cm column using UNOsphere S. Subsequently, scale-up of this newly developed process and conduction of trials at higher scale is also undertaken by applying various scale-up strategies. For these trials, this work categorized various scale-up factors into constant velocity (C V) and constant bed volume per hour (C BVH) scale-up methods. Scale-up trials in bigger diameter (1.5 cm and 2.5 cm) columns with height to diameter ratio (H/D) =19 and <19 were performed. The effects of changing flow velocity and H/D ratio due to the applied scale-up strategy were evaluated and compared with lab performance in terms of resolution vs. lower equivalent to theoretical plate (HETP) relationship. The C BVH method was found to be an efficient method for the scale-up of the developed whey protein resolution system. Also an equation for the prediction of resolution in a scaled column on the basis of a HETP-resolution correlation is presented. It is useful for the prediction of either resolution or bed height in scaled column of WPC or similar biomolecular purification systems.

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Determination of adsorption isotherm parameters for minor whey proteins by gradient elution preparative liquid chromatography

Cited by 13 publications

References 30 publications

Optimization of Lactoperoxidase and Lactoferrin Separation on an Ion-Exchange Chromatography Step

Optimization of Lactoperoxidase and Lactoferrin Separation on an Ion-Exchange Chromatography Step

Optimization of Lactoperoxidase and Lactoferrin Separation on an Ion-Exchange Chromatography Step

Development and Optimization of a Single-Step Cation Chromatographic Whey Protein Fractionation Process: Evaluation and Comparison of Scale-Up Strategies

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