Construction of a highly efficient adsorbent for one-step purification of recombinant proteins: Functionalized cellulose-based monolith fabricated via phase separation method.

Autor: Huang L; School of Pharmacy, Jiangsu University, Zhenjiang 212013, China., Chen G; School of Pharmacy, Jiangsu University, Zhenjiang 212013, China., Zhang G; School of Pharmacy, Jiangsu University, Zhenjiang 212013, China., Fang Y; Department of Geriatrics, Jiangsu University Affiliated People's Hospital, Zhenjiang, China., Zhu W; Department of Bioinformatics and Systems Biology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, Hubei, China., Xin Y; School of Pharmacy, Jiangsu University, Zhenjiang 212013, China. Electronic address: xinyuanrong@ujs.edu.cn.
Jazyk: angličtina
Zdroj: Carbohydrate polymers [Carbohydr Polym] 2024 Jul 01; Vol. 335, pp. 122046. Date of Electronic Publication: 2024 Mar 15.
DOI: 10.1016/j.carbpol.2024.122046
Abstrakt: Currently, purification step in the recombinant protein manufacture is still a great challenge and its cost far outweighs those of the upstream process. In this study, a functionalized cellulose-based monolith was constructed as an efficient affinity adsorbent for one-step purification of recombinant proteins. Firstly, the fundamental cellulose monolith (CE monolith) was fabricated based on thermally induced phase separation, followed by being modified with nitrilotriacetic acid anhydride through esterification to give NCE monolith. After chelating with Ni 2+ , the affinity adsorbent NCE-Ni 2+ monolith was obtained, which was demonstrated to possess a hierarchically porous morphology with a relatively high surface area, porosity and compressive strength. The adsorption behavior of NCE-Ni 2+ monolith towards β2-microglobulin with 6 N-terminus His-tag (His-β2M) was evaluated through batch and fixed-bed column experiments. The results revealed that NCE-Ni 2+ monolith exhibited a relatively fast His-β2M adsorption rate with a maximum adsorption capacity of 329.2 mg/g. The fixed-bed column adsorption implied that NCE-Ni 2+ monolith showed high efficiency for His-β2M adsorption. Finally, NCE-Ni 2+ monolith was demonstrated to have an excellent His-β2M purification ability from E. coli lysate with exceptional reusability. Therefore, the resultant NCE-Ni 2+ monolith had large potential to be used as an efficient adsorbent for recombinant protein purification in practical applications.
Competing Interests: Declaration of competing interest The authors declare they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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