Protein-templated macroporous agarose microspheres as a high-performance chromatographic medium for protein purification

被引:0
作者
Li, Yuqian [1 ]
Chang, Yaoguang [1 ]
Qiu, Junying [3 ]
Yang, Lu [1 ,5 ]
Xue, Changhu [1 ,2 ,4 ]
Zheng, Hongwei [1 ,2 ]
机构
[1] Ocean Univ China, Coll Food Sci & Engn, State Key Lab Marine Food Proc & Safety Control, Qingdao 266404, Peoples R China
[2] Qingdao Inst Marine Bioresources Nutr & Hlth Innov, Qingdao 266109, Peoples R China
[3] Shandong Hailongyuan Biotech Co Ltd, Qingdao 266000, Peoples R China
[4] Qingdao Natl Lab Marine Sci & Technol, Qingdao 266000, Peoples R China
[5] Qingdao Agr Univ, Coll Food Sci & Engn, Qingdao 266109, Shandong, Peoples R China
基金
中国国家自然科学基金;
关键词
Agarose microspheres; Protein separation; Macroporous structure; Epoxy cross-linking; Chromatography; Diethylaminoethyl (DEAE) functionalization; SUPERPOROUS AGAROSE; CROSS-LINKING; ADSORPTION; FABRICATION; SEPARATION; DIFFUSION; HYDROGEL; STRENGTH;
D O I
10.1016/j.carbpol.2025.123266
中图分类号
O69 [应用化学];
学科分类号
081704 ;
摘要
Rapid purification of a targeted protein from a complex medium in advanced biotechnology requires efficient separation platforms. Herein, a new approach for preparing macroporous agarose microspheres (AGs) with enhanced mass transfer stability was developed by a protein-templated strategy. By optimizing the agarose-togelatin ratio (3:1, 3:4, 3:7, and 3:10), interconnected macroporous networks were created, promoting rapid protein diffusion and increasing the accessibility of meso/micropores. Different cross-linking reagents were assessed to enhance the mechanical strength of the AGs, with epichlorohydrin (ECH) demonstrating the highest pressure resistance and epoxy density. To achieve selective protein binding, diethylaminoethyl (DEAE) was directly conjugated to the epoxy-functionalized macroporous AGs (3:4 AGs-ECH and 3:7 AGs-ECH) via reactions involving hydroxyl and chlorine groups. The morphology and chemical composition of the composite AGs were systematically characterized. Benefiting from the macropores, the composites demonstrated significant binding capacities for bovine serum albumin (3:4 AGs-ECH-DEAE: 383.76 mg/g; 3:7 AGs-ECH-DEAE: 369.69 mg/g) as well as bovine hemoglobin (3:4 AGs-ECH-DEAE: 363.94 mg/g; 3:7 AGs-ECH-DEAE: 237.58 mg/g). Besides, the prepared 3: 4 AGs-ECH-DEAE and 3: 7 AGs-ECH-DEAE media exhibited ideal separation effects in chromatographic tests compared to DEAE Sepharose Fast Flow, highlighting their potential as effective chromatographic media for the swift separation and purification of proteins.
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页数:16
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