A simple and efficient purification platform for monoclonal antibody production based on chromatin-directed cell culture clarification integrated with precipitation and void-exclusion anion exchange chromatography | |
Chen, Quan1; Latiff, Sarah Maria Abdul2; Toh, Phyllicia2; Peng, Xinying1; Hoi, Aina2; Xian, Mo1; Zhang, Haibo1; Nian, Rui1; Zhang, Wei2; Gagnon, Pete2 | |
2016-10-20 | |
发表期刊 | JOURNAL OF BIOTECHNOLOGY |
卷号 | 236页码:128-140 |
摘要 | Protein A affinity chromatography, featured by its robustness and high-specificity, is still dominant as a first capture step for the purification of immunoglobulin G monoclonal antibodies (IgG mAbs). However, the material and operational costs of protein A are universally recognized as high, and its productivity is also limited as column mode. In order to overcome these limitations, industry is increasingly considering the use of non-protein A-based processes for IgG purification. In this study, sodium citrate precipitation (SCP) was developed as the primary purification step, and chromatin-directed cell culture clarification was demonstrated to significantly elevate the purification capability. Additional 0.05% (w/v) of Tween 20 was shown to effectively reduce the residual free antibody light chain (LC) during precipitation. The resuspended IgG was further polished by void-exclusion anion exchange chromatography (VEAX), which supported protein loading without buffer adjustment. The non-histone host cell protein (nh-HCP) content in the final product was about 5 ppm and histone HCP below limit of detection (LOD). DNA was reduced to less than 1 ppb, and aggregates/free LC less than 0.1%. The overall IgG recovery was 87.2%. A simple and efficient purification platform with only one-column step was therefore established, providing a more promising alternative to the current prevailing protein A-based purification platforms. (C) 2016 Elsevier B.V. All rights reserved. |
文章类型 | Article |
关键词 | Precipitation Monoclonal Antibody Purification Chromatin-directed Clarification Light Chain Surfactant |
WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
DOI | 10.1016/j.jbiotec.2016.08.014 |
关键词[WOS] | A AFFINITY-CHROMATOGRAPHY ; MIXED-MODE CHROMATOGRAPHY ; IMMUNOGLOBULIN-G ; BIOSEPARATION TECHNIQUE ; HUMAN-ALBUMIN ; FUTURE ; CAPTURE ; IGG ; MICROFILTRATION ; IDENTIFICATION |
收录类别 | SCI |
语种 | 英语 |
WOS研究方向 | Biotechnology & Applied Microbiology |
项目资助者 | QIBEBT (Qingdao Institute of Bioenergy and Bioprocess Technology) Start-up Fund(Y571061905) ; Biomedical Research Council of A*STAR ; Exploit Technologies Pte Ltd of Singapore(ETPL/12-R15GAP-0009) |
WOS类目 | Biotechnology & Applied Microbiology |
WOS记录号 | WOS:000384852600013 |
引用统计 | |
文献类型 | 期刊论文 |
条目标识符 | http://ir.qibebt.ac.cn/handle/337004/9082 |
专题 | 生物基材料组群 |
作者单位 | 1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Key Lab Biobased Mat, 189 Songling Rd, Qingdao 266101, Peoples R China 2.Agcy Sci Technol & Res, Bioproc Technol Inst, 20 Biopolis Way, Singapore 138668, Singapore |
推荐引用方式 GB/T 7714 | Chen, Quan,Latiff, Sarah Maria Abdul,Toh, Phyllicia,et al. A simple and efficient purification platform for monoclonal antibody production based on chromatin-directed cell culture clarification integrated with precipitation and void-exclusion anion exchange chromatography[J]. JOURNAL OF BIOTECHNOLOGY,2016,236:128-140. |
APA | Chen, Quan.,Latiff, Sarah Maria Abdul.,Toh, Phyllicia.,Peng, Xinying.,Hoi, Aina.,...&Gagnon, Pete.(2016).A simple and efficient purification platform for monoclonal antibody production based on chromatin-directed cell culture clarification integrated with precipitation and void-exclusion anion exchange chromatography.JOURNAL OF BIOTECHNOLOGY,236,128-140. |
MLA | Chen, Quan,et al."A simple and efficient purification platform for monoclonal antibody production based on chromatin-directed cell culture clarification integrated with precipitation and void-exclusion anion exchange chromatography".JOURNAL OF BIOTECHNOLOGY 236(2016):128-140. |
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