STERIC MASS ACTION MODEL FOR LACTOFERRIN ADSORPTION IN CRYOGEL WITH IMMOBILIZED COPPER IONS

B M A Carvalho,L M Carvalho,G G P Carvalho,L A Minim,W F Silva Júnior

doi:10.1590/0104-6632.20160331s20140112

Abstract

Parameters of equilibrium adsorption obtained from experiments using immobilized metal affinity chromatography (IMAC) were used to evaluate the applicability of the steric mass-action (SMA) model to describe the adsorption of lactoferrin to cryogel resin under different conditions. The adsorption of lactoferrin on continuous supermacroporous cryogel with immobilized Cu2+ ions was evaluated in batch adsorption experiments at different pH (6-8) and temperature (293-313 K) values. Estimated values of the equilibrium constant (K) and characteristic number of binding sites (n) showed that these parameters decreased with increasing ionic strength, pH and temperature, while the nonlinear parameter, the steric factor (σ), increased with increasing ionic strength and temperature. Expressions correlating these parameters with pH, ionic strength and temperature were then derived.

Highlights

The recent requirements of high purity proteins, and the need of reducing costs of downstream processes have stimulated the development of more efficient and cheaper separation techniques (Frerick et al, 2008)
Parameters of equilibrium adsorption obtained from experiments using immobilized metal affinity chromatography (IMAC) were used to evaluate the applicability of the steric mass-action (SMA) model to describe the adsorption of lactoferrin to cryogel resin under different conditions
Recent studies have used the SMA model to describe protein adsorption equilibrium and the results have shown its efficiency for predicting the non-linear adsorption behavior of proteins (Chen et al, 2006; Barz et al, 2010)

Summary

Introduction

The recent requirements of high purity proteins (both natural and recombinant), and the need of reducing costs of downstream processes have stimulated the development of more efficient and cheaper separation techniques (Frerick et al, 2008). It has been shown that Immobilized Metal Affinity Chromatography (IMAC) meets these requirements (Carvalho et al, 2014; Wang et al, 2008; Cheung et al, 2012) This technology is based on the chemical affinity displayed by certain groups of amino acid residues present on the surface of proteins (e.g., the IMAC has been reported as a promissing technology for purification of proteins with an “N-terminal metal biding tag” (e.g., histidine tail and NT1A) (Puri et al, 2010; Petzold et al, 2014; Cheung et al, 2012), including on a large scale (Gaberc-Porekar and Menart, 2005), due to its low cost and the high purity of the eluate, even in a single step process (Puri et al, 2010; Carvalho et al, 2014). Its use on an industrial scale has still been only slightly explored

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Conclusion