title: A mixed-mode resin with tryptamine ligand for human serum albumin separation.
abstract: Mixed-mode chromatography (MMC) is a new technology that uses specially-designed ligands to improve the adsorption selectivity with multimodal protein-ligand interactions for protein separation. A new MMC resin TA-B-6FF with tryptamine as the functional ligand was prepared and used for human serum albumin (HSA) separation. Adsorption equilibria of plasma-derived HSA (pHSA) were investigated and compared with a commercial tryptophan-based resin (MX-Trp-650m), and the influence of pH and salt addition was studied. The results showed that weak acidic conditions (pH 5.0-7.0) were favorable for HSA adsorption. The maximum adsorption capacity of TA-B-6FF was 141.33mg/g at pH 5.0, which was two times higher than that of MX-Trp-650m. TA-B-6FF also showed better salt-tolerance than MX-Trp-650m. Moreover, TA-B-6FF was used to separate recombinant HSA (rHSA) from Pichia pastoris culture broth. The results indicated that rHSA could be directly captured by TA-B-6FF without dilution or pH adjustment. High purity (87.75%) of rHSA monomer could be obtained with a recovery of 98.53% through two-step elution process. Total content of rHSA monomer and degraded fragment was 99.75%, the removal of host cell proteins reached about 90%. The results demonstrate that new TA-B-6FF resin has a great potential for rHSA purification directly from the complex fermentation broth.
authors.affiliation: Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Qi-Ci
authors.initials: QC
authors.lastname: Wu
authors.affiliation: Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China. Electronic address: lindq@zju.edu.cn.
authors.forename: Dong-Qiang
authors.initials: DQ
authors.lastname: Lin
authors.affiliation: Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Shi
authors.affiliation: Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Qi-Lei
authors.initials: QL
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Shan-Jing
authors.initials: SJ
authors.lastname: Yao
mesh_terms: D000327:Adsorption; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002845:Chromatography / Q000379:methods*; D003470:Culture Media / Q000737:chemistry; D006801:Humans; D007475:Ion Exchange Resins; D008024:Ligands; D010843:Pichia / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry; D014363:Tryptamines / Q000737:chemistry*; D014364:Tryptophan / Q000737:chemistry
chemical_list: D003470:Culture Media; D007475:Ion Exchange Resins; D008024:Ligands; D012709:Serum Albumin; D014363:Tryptamines; C030820:tryptamine; D012965:Sodium Chloride; D014364:Tryptophan
keywords: Adsorption; Human serum albumin; Mixed-mode chromatography; Pichia pastoris; Tryptamine