Preparation and evaluation of mixed-mode resins with tryptophan analogues as functional ligands for human serum albumin separation

doi:10.1016/j.cjche.2016.12.010

摘要/Abstract

摘要： Five tryptophan analogues with a hydrophobic indole ring and an amino group on each molecule were used as functional ligands of mixed-mode resins for human serum albumin (HSA) purification. Their adsorption performance was evaluated and the effects of pH and salt addition on HSA adsorption were studied. The resins prepared showed typical pH-dependent adsorption and the highest adsorption capacity and affinity were found at pH 5.0 for all the resins tested. The saturated adsorption capacity was 138.02 mg·g^-1 with the tryptaminefunctionalized resin, which significantly decreased at pH below 4.0 due to electrostatic repulsion between ligands and HSA. Moreover, the addition of NaCl or (NH₄)₂SO₄ in media reduced HSA adsorption capacity, although the two salts showed different affecting profiles. The tryptamine-functionalized resin showed the best salt-tolerant performance, and its high adsorption capacity was maintained under high salt concentrations. In addition, the five resins prepared showed good adsorption selectivity for recombinant HSA from Pichia pastoris broth. Molecular docking results between tryptamine and HSA indicated that tryptamine was favorable to bind on Site Ⅱ (indole-binding site) of HSA.

关键词: Adsorption, Human serum albumin, Mixed-mode chromatography, Tryptamine, Tryptophan analogue

Abstract: Five tryptophan analogues with a hydrophobic indole ring and an amino group on each molecule were used as functional ligands of mixed-mode resins for human serum albumin (HSA) purification. Their adsorption performance was evaluated and the effects of pH and salt addition on HSA adsorption were studied. The resins prepared showed typical pH-dependent adsorption and the highest adsorption capacity and affinity were found at pH 5.0 for all the resins tested. The saturated adsorption capacity was 138.02 mg·g^-1 with the tryptaminefunctionalized resin, which significantly decreased at pH below 4.0 due to electrostatic repulsion between ligands and HSA. Moreover, the addition of NaCl or (NH₄)₂SO₄ in media reduced HSA adsorption capacity, although the two salts showed different affecting profiles. The tryptamine-functionalized resin showed the best salt-tolerant performance, and its high adsorption capacity was maintained under high salt concentrations. In addition, the five resins prepared showed good adsorption selectivity for recombinant HSA from Pichia pastoris broth. Molecular docking results between tryptamine and HSA indicated that tryptamine was favorable to bind on Site Ⅱ (indole-binding site) of HSA.

Key words: Adsorption, Human serum albumin, Mixed-mode chromatography, Tryptamine, Tryptophan analogue

Qici Wu, Qilei Zhang, Shiwen Xu, Cheng ong Ge, Shanjing Yao, Dongqiang Lin. Preparation and evaluation of mixed-mode resins with tryptophan analogues as functional ligands for human serum albumin separation[J]. , 2017, 25(7): 898-905.

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