Investigation of protein retention and selectivity in HIC systems using quantitative structure retention relationship models

Ladiwala, Asif; Xia, Fang; Luo, Qijun; Breneman, Curt M.; Cramer, Steven M.

doi:10.1002/bit.20771

Cited by 69 publications

(62 citation statements)

References 41 publications

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“…Prediction of HIC adsorption is an active area of research with many recent publications focusing on relating the retention to crystal structure (Lienqueo et al, 2002;Mahn et al, 2004;Salgado et al, 2006a,b), descriptors that are derived from the crystal structure (Ladiwala et al, 2006), or the amino acid sequence (Salgado et al, 2005). We have observed that, for wild-type a-lactalbumin, there may be some unfolding on the surface with little impact on the observed retention, and predictive methods that assume native conformation on the surface might be applied successfully.…”

Section: Adsorption Of Disulfide Bond-reduced Variantsmentioning

confidence: 99%

Protein instability during HIC: Evidence of unfolding reversibility, and apparent adsorption strength of disulfide bond‐reduced α‐lactalbumin variants

Deitcher

Xiao

O’Connell

et al. 2009

Biotech & Bioengineering

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A two-conformation, four-state model has been proposed to describe protein adsorption and unfolding behavior on hydrophobic interaction chromatography (HIC) resins. In this work, we build upon previous study and application of a four-state model to the effect of salt concentration on the adsorption and unfolding of the model protein alpha-lactalbumin in HIC. Contributions to the apparent adsorption strength of the wild-type protein from native and unfolded conformations, obtained using a deuterium labeling technique, reveal the free energy change and kinetics of unfolding on the resin, and demonstrate that surface unfolding is reversible. Additionally, variants of alpha-lactalbumin in which one of the disulfide bonds is reduced were synthesized to examine the effects of conformational stability on apparent retention. Below the melting temperatures of the wild-type protein and variants, reduction of a single disulfide bond significantly increases the apparent adsorption strength (approximately 6-8 kJ/mol) due to increased instability of the protein. Finally, the four-state model is used to accurately predict the apparent adsorption strength of a disulfide bond-reduced variant.

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Section: Adsorption Of Disulfide Bond-reduced Variantsmentioning

confidence: 99%

Protein instability during HIC: Evidence of unfolding reversibility, and apparent adsorption strength of disulfide bond‐reduced α‐lactalbumin variants

Deitcher

Xiao

O’Connell

et al. 2009

Biotech & Bioengineering

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“…Since this technique is labor intensive and requires significant effort to determine the conditions for optimal separation, a computational model for predicting the behavior of proteins in HIC systems is a valuable tool. Previously, Ladiwala et al [95] had developed a quantitative structureretention relationship (QSRR) model for protein retention time in HIC systems based on a support vector machine (SVM) algorithm using three different categories of descriptors for the proteins, including a novel set of protein hydrophobicity descriptors. Costache et al [92] reconsidered the experimental data of Ladiwala [95] and used the ''Biomaterials Store TM '' and a different approach to predict the protein retention time based on a hybrid DT and ANN model and just traditional 2-D and 3-D descriptors based on protein structures.…”

Section: Online Databases and Computational Toolsmentioning

confidence: 99%

Computational Methods for the Development of Polymeric Biomaterials

Costache¹,

Ghosh²,

Knight³

et al. 2010

Adv Eng Mater

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“…On the other hand, the protein's average surface hydrophobicity (Lienqueo et al, 2003), local hydrophobic distribution (Mahn et al, 2005), and statistical description of protein surface amino acid distribution (Salgado et al, 2005) have been used as a measure to predict the retention in HIC systems. Additionally, a quantitative structure-retention relationship (QSRR) model, based on a support vector machine (SVM), has been proposed for evaluating the effects of stationary phase resin chemistry and protein physicochemical properties on protein binding affinity in HIC (Ladiwala et al, 2006). Another mathematical model, such as the Plate Model, is mainly used for predicting retention times and elution curves.…”

Section: Introductionmentioning

confidence: 99%

Optimization of hydrophobic interaction chromatography using a mathematical model of elution curves of a protein mixture

Lienqueo

Shene

Asenjo

2008

J of Molecular Recognition

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This paper describes a methodology for optimizing performance of hydrophobic interaction chromatography (HIC) for protein mixtures in which Rate Model simulations and evaluation of cost function are used. The system under study was HIC of a two-protein mixture (alpha-chymotrypsin and alpha-amylase), carried out with different conditions (gradient steepness, salt, concentration, volume of sample, pH, type of matrix, and flow rates). Parameters in the rate model were obtained from different sources. Mass transfer parameters (Reynolds, Peclet, and Biot numbers) were calculated using empirical correlations. Under the experimental conditions Re number was small (0.23) and axial dispersion negligible (PeL>300). Mass transfer was controlled by intraparticle diffusion (Bi>50). The model assumes that equilibrium constant (b) in the Langmuir isotherm was salt concentration (I) dependent [b(I)]. Parameters in the relationship for b(I) were estimated from experimental single protein elution curves and used to simulate protein mixtures. Rate model simulations showed that if protein sample load to the column was below 1 mg, displacement effects were negligible; in other cases protein interactions would limit the proposed mathematical description of HIC. The calibrated Rate Model successfully predicted elution curves of the protein mixture with deviations lower than 6x10(-4) absorbance units. The model was also able to predict that the Butyl Sepharose--NaCl 4 M system allowed to obtain the highest resolution (>1) for the two-protein mixture evaluated. The cost function built for optimizing the performance of HIC considers yield, purity, concentration, and the time needed to accomplish the separation. This function contains two types of parameters that have to be determined. The ones dependent on the HIC system and process conditions were obtained from simulations of elution curves of the two-protein mixture, by the calibrated Rate Model. The other parameters are dependent on characteristic and quality of the protein product; these were assumed for illustration purpose. Minimization of the cost function allows determination of flow rate, gradient steepness, and the fraction of eluted peak that has to be collected. Novelty of the present work is in showing how parameters in the fundamentally based Rate Model for HIC can be calibrated and how simulations can be used for the optimization of process conditions for the separation of a protein mixture.

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Investigation of protein retention and selectivity in HIC systems using quantitative structure retention relationship models

Cited by 69 publications

References 41 publications

Protein instability during HIC: Evidence of unfolding reversibility, and apparent adsorption strength of disulfide bond‐reduced α‐lactalbumin variants

Protein instability during HIC: Evidence of unfolding reversibility, and apparent adsorption strength of disulfide bond‐reduced α‐lactalbumin variants

Computational Methods for the Development of Polymeric Biomaterials

Optimization of hydrophobic interaction chromatography using a mathematical model of elution curves of a protein mixture

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