适应高滴度抗体制备的抗体捕集纯化技术研究进展

doi:10.16085/j.issn.1000-6613.2018-1115

化工进展 ›› 2019, Vol. 38 ›› Issue (01): 576-585.DOI: 10.16085/j.issn.1000-6613.2018-1115

适应高滴度抗体制备的抗体捕集纯化技术研究进展

史清洪^1,²(),孙彦^1,²()

1. 天津大学化工学院，天津 300350
2. 系统生物工程教育部重点实验室，天津 300350

收稿日期:2018-05-30 修回日期:2018-08-24 出版日期:2019-01-05 发布日期:2019-01-05
通讯作者: 孙彦
作者简介:史清洪（1968—），男，工学博士，教授，博士生导师，研究方向为生物分离工程。E-mail：<email>qhshi@tju.edu.cn</email>。|孙彦，教授，博士生导师，研究方向为生物分子工程。E-mail：<email>ysun@tju.edu.cn</email>。
基金资助:
国家自然科学基金面上项目(21476166, 21878221);国家自然科学基金创新群体项目(21621004);国家自然科学基金重点项目(21236005);国家自然科学基金面上项目（21476166, 21878221）；国家自然科学基金创新群体项目（21621004）；国家自然科学基金重点项目（21236005）。

Antibody capture technologies for high-titer antibody production

Qinghong SHI^1,²(),Yan SUN^1,²()

1. School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350，China
2. Key Laboratory of Systems Bioengineering Ministry of Education, Tianjin 300350, China

Received:2018-05-30 Revised:2018-08-24 Online:2019-01-05 Published:2019-01-05
Contact: Yan SUN

摘要/Abstract

摘要：

抗体捕集纯化是单克隆抗体和Fc融合蛋白药物制备过程的关键步骤。但相较于大规模动物细胞培养技术的迅猛发展，抗体的捕集纯化已经成为了制约单抗药物生产的主要“瓶颈”。本文回顾了蛋白A亲和层析和阳离子交换层析等当前抗体药物工业生产过程主要捕集纯化技术的发展现状和应用情况，介绍了近年来亲和肽层析和混合模式吸附层析等新型层析分离技术的发展以及膨胀床吸附和多柱串联吸附等过程集成方法在提高抗体捕集效率方面所展现的良好前景。在此基础上，指出了影响抗体捕集纯化的主要因素以及各技术发展中存在的问题，展望抗体捕集纯化技术的发展方向。

关键词: 抗体, 捕集纯化, 蛋白A层析, 阳离子交换层析, 亲和肽层析, 混合模式层析, 膨胀床吸附, 多柱串联吸附

Abstract:

Product capture from a cell culture broth is a key step in the production of antibodies and Fc-fusion proteins. With a rapid increase of large-scale animal cell culture techniques, capture techniques in antibody production have become the key technical bottleneck in monoclonal antibody productions. In this paper, core antibody capture methods, including protein A chromatography and cation exchange chromatography, have been reviewed in term of their history and applications. At the same time, peptide affinity chromatography, mixed-mode or multimodal chromatography, expanded-bed adsorption and sequential multi-column adsorption have been introduced to exhibit their great potentials in high-efficient antibody captures. Moreover, the factors affecting the capture and purification processes were analyzed while the problems that need to be solved to improve the technologies were discussed. Finally, perspectives into the development of antibody captures were discussed with focusing on the technological aspects of the chromatographic methods.

Key words: antibody, capture techniques, protein A chromatography, cation exchange chromatography, peptide affinity chromatography, mixed-mode chromatography, expanded-bed adsorption, sequential multi-column adsorption

中图分类号:

TQ028.8

史清洪, 孙彦. 适应高滴度抗体制备的抗体捕集纯化技术研究进展[J]. 化工进展, 2019, 38(01): 576-585.

Qinghong SHI, Yan SUN. Antibody capture technologies for high-titer antibody production[J]. Chemical Industry and Engineering Progress, 2019, 38(01): 576-585.

图/表 2

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介质名称	基质材料	配基种类	配基密度 /mg·mL^-1	介质性能				参考文献
介质名称	基质材料	配基种类	配基密度 /mg·mL^-1	饱和吸附容量/mg·mL^-1	解离常数 /mg·mL^-1	动态吸附容量 /mg·mL^-1		参考文献
GE Healthcare
rProtein A Sepharose Fast Flow	交联琼脂糖		4～6	55.1±1.5	0.037±0.007			[18]
Protein A Sepharose 4 Fast Flow	交联琼脂糖		4～6	61.6±1.7	0.11±0.09			[18]
MabSelect	高度交联琼脂糖	重组蛋白A	n.a.	60.6±1.3	0.120±0.010	约30		[31]
MabSelect SuRe	高度交联琼脂糖	碱耐受性突变蛋白A	n.a.	61.2±2.1	0.115±0.017	约30		[31]
MabSelect Xtra	高度交联琼脂糖	重组蛋白A	n.a.	68.3±2.0	0.088±0.011	约40		[31]
MabSelect PrismA	高度交联琼脂糖	碱耐受性突变蛋白A	n.a.	99.9	—	>80
Millipore
ProSep-vA Ultra	可控多孔玻璃材料	天然蛋白A	n.a.	62.8±1.8	0.116±0.014	35		[31]
ProSep-vA High capacity	可控多孔玻璃材料		n.a.	>40	—	>20
ProSep-vA Ultra Plus	可控多孔玻璃材料	重组蛋白A	n.a.	>67	—	>50
Eshmuno A resin	亲水聚乙烯醚	重组C结构域	n.a.	—	—	40～55
PerSecptive Biosystem
POROS 50 A	交联聚（苯乙烯-二乙烯基苯）	重组蛋白A	n.a.	25	—		17.5	^[37]	[32]
POROS MabCapture A	交联聚（苯乙烯-二乙烯基苯）	重组蛋白A	n.a.	—	—		>45

介质名称	基质材料	配基种类	配基密度 /mg·mL^-1	介质性能				参考文献
介质名称	基质材料	配基种类	配基密度 /mg·mL^-1	饱和吸附容量/mg·mL^-1	解离常数 /mg·mL^-1	动态吸附容量 /mg·mL^-1		参考文献
GE Healthcare
rProtein A Sepharose Fast Flow	交联琼脂糖		4～6	55.1±1.5	0.037±0.007			[18]
Protein A Sepharose 4 Fast Flow	交联琼脂糖		4～6	61.6±1.7	0.11±0.09			[18]
MabSelect	高度交联琼脂糖	重组蛋白A	n.a.	60.6±1.3	0.120±0.010	约30		[31]
MabSelect SuRe	高度交联琼脂糖	碱耐受性突变蛋白A	n.a.	61.2±2.1	0.115±0.017	约30		[31]
MabSelect Xtra	高度交联琼脂糖	重组蛋白A	n.a.	68.3±2.0	0.088±0.011	约40		[31]
MabSelect PrismA	高度交联琼脂糖	碱耐受性突变蛋白A	n.a.	99.9	—	>80
Millipore
ProSep-vA Ultra	可控多孔玻璃材料	天然蛋白A	n.a.	62.8±1.8	0.116±0.014	35		[31]
ProSep-vA High capacity	可控多孔玻璃材料		n.a.	>40	—	>20
ProSep-vA Ultra Plus	可控多孔玻璃材料	重组蛋白A	n.a.	>67	—	>50
Eshmuno A resin	亲水聚乙烯醚	重组C结构域	n.a.	—	—	40～55
PerSecptive Biosystem
POROS 50 A	交联聚（苯乙烯-二乙烯基苯）	重组蛋白A	n.a.	25	—		17.5	^[37]	[32]
POROS MabCapture A	交联聚（苯乙烯-二乙烯基苯）	重组蛋白A	n.a.	—	—		>45

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适应高滴度抗体制备的抗体捕集纯化技术研究进展

Antibody capture technologies for high-titer antibody production

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