Preparative affinity chromatography of proteins

Narayanan, Sunanda R.

doi:10.1016/0021-9673(94)80018-9

Cited by 83 publications

(37 citation statements)

References 88 publications

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“…There are now examples of at least 30 different dyes for the puri®cation of a great variety of proteins ranging through recombinant polypeptide hormones, blood proteins, and most classes of enzymes [2]. Dyes have often replaced nucleotides as af®nity ligand in large-scale puri®cation of biomolecules [3].…”

Section: Introductionmentioning

confidence: 99%

Adsorption-desorption of BSA to highly substituted dye-ligand adsorbent: quantitative study of the effect of ionic strength

Gan

Sun

1997

Bioprocess Engineering

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Cibacron Blue 3GA was immobilized on Sepharose CL-6B to obtain a highly substituted dye-ligand adsorbent which dye concentration was 17.4 lmol dye per gram wet gel. This adsorbent had a highly binding capacity for bovine serum albumin (BSA). The effects of ionic strength on the adsorption and desorption of BSA to the adsorbent were studied. Adsorption isotherms were expressed by the Langmuir model. The quantitative relationships between the model parameters and the ionic strength were obtained. The desorptions were performed by adding salt to the BSA solutions in which adsorption equilibria had been reached. Adding salt to the solution resulted in the desorption of the bound protein. It was found that the isotherm obtained from the desorption experiments agreed well to the isotherm obtained from the adsorption experiments at the same ionic strength. The result demonstrated that the adsorption of BSA to the highly substituted adsorbent was reversible.List of symbols C mg cm A3 liquid phase concentration of BSA at equilibrium C 0 mg cm A3 the concentration of the initial protein solution C 1 mg cm A3 liquid phase concentration of BSA at ®rst equilibrium C 2 mg cm A3 liquid phase concentration of BSA after desorption C s mol dm A3 the salt concentration K d mg cm A3 dissociation constant in Langmuir model q mg g A1 adsorbed density of BSA at equilibrium q 1 mg g A1 adsorbed BSA density at ®rst equilibrium q 2 mg g A1 adsorbed BSA density after desorption q m mg g A1 Langmuir parameter, adsorption capacity for BSA V 0 cm A3 volume of the initial protein solution V 1 cm A3 withdrawn volume of protein solution for earlier adsorption determination V 2 cm A3 added volume of the buffer solution for desorption W g wet weight of the adsorbent 1 Introduction Dye-ligand adsorbents for protein puri®cations have been developed extensively because they offer the chance of simple and rapid isolations of proteins, typically 20 to 80 fold puri®cation with about 80% recovery from a crude extract without prior treatment [1]. There are now examples of at least 30 different dyes for the puri®cation of a great variety of proteins ranging through recombinant polypeptide hormones, blood proteins, and most classes of enzymes [2]. Dyes have often replaced nucleotides as af®nity ligand in large-scale puri®cation of biomolecules [3]. There are several examples of large-scale puri®cation of proteins using dye ligand af®nity chromatography, such as glycerokinase, ATP: AMP phospho-transferase [4,5]. More recent alternative exploitations of dye-protein interactions such as af®nity cross-¯ow ®ltration, af®nity precipitation and af®nity extraction also play an important role in such isolations [6±9]. Among all of the dyes, the mostly utilized one is a reactive dye, Cibacron Blue F3GA (Fig. 1), which is a mixture of two isomers [3, 10, 11]. There are many factors affecting the interactions between a protein and an immobilized dye, such as immobilized dye density, ionic strength, temperature, pH and buffer composition [1,8,9,12]. It was found that th...

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Section: Introductionmentioning

confidence: 99%

Adsorption-desorption of BSA to highly substituted dye-ligand adsorbent: quantitative study of the effect of ionic strength

Gan

Sun

1997

Bioprocess Engineering

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“…[16][17] We suppose that the bond between b-glucosylamidine and b-glucoside may be hydrogen bond, static electronic bond, hydrophobic reaction, van der Waals force or some other kind of weak bond. The mechanism is still under research.…”

Section: Separation Function Of the Molecular Recognition Bionic Chromentioning

confidence: 99%

Preparation and Evaluation of a Molecular Recognition Bionic Solid Phase Extraction Column for Separation of Glucosides

Tang

Cai

Gao

et al. 2010

J Chinese Chemical Soc

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A molecular recognition bionic solid phase extraction (SPE) column for separation of glucosides has been prepared using a positively charged b-glucosylamidine as the ligand in which a glycon moiety is connected via an N-glycoside linkage. b-Glucosylamidine, highly potent and selective inhibitors of b-glycosidase, is immobilized through a one-step synthesize procedure involving the addition of b-glucosylamine and 2-iminothiolane. HCl simultaneously to a matrix modified with maleimido groups via an appropriate spacer to give a molecular recognition absorbent for b-glucosides. N-octyl-b-D-glucopyranoside and b-D-galactopyranoside or a-D-mannopyranoside was directly chromatographed through the bionic chromatographic column, resulting in a much stronger retention of b-D-glucopyranoside than b-D-galactopyranoside and a-D-mannopyranoside. The retained glucopyranoside could only be eluted by glucose solution. This indicates that the binding of the glucoside was of specific nature that corresponds to the glycon substrate specificity of the glucoside. The ease of preparation and the selective nature of the molecular recognition bionic chromatography should promise a large-scale preparation of the molecular recognition adsorbent for the purification and removal of glucosides according to their glycon substrate specificity.

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“…To improve this behavior, the new types of dispersed sorbents were suggested over the past ten years. Among these are nonporous [21,22], micropellicular [13, 19, 23 -25], or the gigaporous structures found in the so-called "perfusion" [14 -17] and the "gel in a shell" [20] sorbent beads, but also a number of beads with improved accessibility of the active ligands [18,26], as well as some new and original designs of the separation modules and methods [28].…”

Section: Dispersed Particlesmentioning

confidence: 99%

Chromatographic Reactors Based on Biological Activity

Podgornik

Tennikova

2002

Advances in Biochemical Engineering/Biotechnology

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In the last decade there were many papers published on the study of enzyme catalyzed reactions performed in so-called chromatographic reactors. The attractive feature of such systems is that during the course of the reaction the compounds are already separated, which can drive the reaction beyond the thermodynamic equilibrium as well as remove putative inhibitors. In this chapter, an overview of such chromatographic bioreactor systems is given. Besides, some immobilization techniques to improve enzyme activity are discussed together with modern chromatographic supports with improved hydrodynamic characteristics to be used in this context.

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Preparative affinity chromatography of proteins

Cited by 83 publications

References 88 publications

Adsorption-desorption of BSA to highly substituted dye-ligand adsorbent: quantitative study of the effect of ionic strength

Adsorption-desorption of BSA to highly substituted dye-ligand adsorbent: quantitative study of the effect of ionic strength

Preparation and Evaluation of a Molecular Recognition Bionic Solid Phase Extraction Column for Separation of Glucosides

Chromatographic Reactors Based on Biological Activity

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