Towards continuous mAb purification: Clearance of host cell proteins from CHO cell culture harvests via “flow‐through affinity chromatography” using peptide‐based adsorbents
Towards continuous mAb purification: Clearance of host cell proteins from CHO cell culture harvests via “flow‐through affinity chromatography” using peptide‐based adsorbents
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实现连续 mAb 纯化：使用基于肽的吸附剂，通过“流动”和“亲和层析”从 CHO 细胞培养物收获物中清除宿主细胞蛋白
DOI：
10.1002/bit.28096
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发表时间：
2022
影响因子：
3.8
通讯作者：
Yigzaw, Yinges
中科院分区：
文献类型：
--
作者：
Sripada, Sobhana Alekhya;Chu, Wenning;Williams, Taufika Islam;Teten, Matthew A.;Mosley, Brian J.;Carbonell, Ruben G.;Lenhoff, Abraham M.;Cramer, Steven M.;Bill, Jerome;Yigzaw, Yinges
单克隆抗体（mAb）生产中先进分析技术的发展凸显了与宿主细胞蛋白（HCP）清除相关的挑战。特别值得关注的是去除“持久性”HCP，包括免疫原性和mAb降解蛋白质，这些蛋白质从蛋白A树脂中共沉淀，可以逃避抛光步骤。为了应对这一挑战，我们引入了一系列肽配体，这些肽配体靶向中国仓鼠卵巢（CHO）细胞培养液中的HCP，并能够通过流通亲和色谱法纯化mAb。本研究描述了它们与LigaGuard™的整合，LigaGuard ™是一种亲和吸附剂，其平衡结合容量为约30 mg HCP/mL树脂，在1 min和2 min停留时间下的动态容量分别高达16 mg/mL和22 mg/mL。当针对具有不同mAb和HCP滴度和性质的细胞培养收获物进行评价时，LigaGuard™提供了高HCP清除率，对数去除值（LRV）高达1.5，mAb产率高于90%。流出物的蛋白质组学分析证实去除了高风险HCP，包括组织蛋白酶、组蛋白、谷胱甘肽-S转移酶和脂蛋白脂肪酶。最后，将用于HCP去除的LigaGuard™与用于产物捕获的亲和吸附剂组合提供85%的总体mAb产率，并且HCP和DNA LRV> 4。
The growth of advanced analytics in manufacturing monoclonal antibodies (mAbs) has highlighted the challenges associated with the clearance of host cell proteins (HCPs). Of special concern is the removal of “persistent” HCPs, including immunogenic and mAb‐degrading proteins, that co‐elute from the Protein A resin and can escape the polishing steps. Responding to this challenge, we introduced an ensemble of peptide ligands that target the HCPs in Chinese hamster ovary (CHO) cell culture fluids and enable mAb purification via flow‐through affinity chromatography. This study describes their integration into LigaGuard™, an affinity adsorbent featuring an equilibrium binding capacity of ~30 mg of HCPs per mL of resin as well as dynamic capacities up to 16 and 22 mg/ml at 1‐ and 2‐min residence times, respectively. When evaluated against cell culture harvests with different mAb and HCP titers and properties, LigaGuard™ afforded high HCP clearance, with logarithmic removal values (LRVs) up to 1.5, and mAb yield above 90%. Proteomic analysis of the effluents confirmed the removal of high‐risk HCPs, including cathepsins, histones, glutathione‐S transferase, and lipoprotein lipases. Finally, combining LigaGuard™ for HCP removal with affinity adsorbents for product capture afforded a global mAb yield of 85%, and HCP and DNA LRVs > 4.
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开发用于从细胞培养物生产收获物中靶向捕获宿主细胞蛋白的肽配体。
影响因子：
--
作者：
R. A. Lavoie;T. Williams;R. K. Blackburn;R. Carbonell;S. Menegatti
通讯作者：
S. Menegatti
影响因子：
--
作者：
Li X;Wang F;Li H;Richardson DD;Roush DJ
通讯作者：
Roush DJ
DOI：
10.1039/c7an01469d
发表时间：
2018-01-15
期刊：
The Analyst
影响因子：
--
作者：
Zhang C ;Rodriguez E ;Bi C ;Zheng X ;Suresh D ;Suh K ;Li Z ;Elsebaei F ;Hage DS
通讯作者：
Hage DS
影响因子：
5.6
作者：
Lavoie, R. Ashton;di Fazio, Alice;Menegatti, Stefano
通讯作者：
Menegatti, Stefano
影响因子：
19
作者：
R. Prodromou;Kevin N. Day;Sahand Saberi-Bosari;John D Schneible;Matthew D. Mabe;A. San Miguel;M. Daniele;V. Pozdin;S. Menegatti
通讯作者：
R. Prodromou;Kevin N. Day;Sahand Saberi-Bosari;John D Schneible;Matthew D. Mabe;A. San Miguel;M. Daniele;V. Pozdin;S. Menegatti