添加SNS能显著提高CHT填料使用寿命的研究

doi:10.13523/j.cb.20170112

中国生物工程杂志

2017, Vol. 37

Issue (1): 81-88 DOI: 10.13523/j.cb.20170112

技术与方法

添加SNS能显著提高CHT填料使用寿命的研究

邓义熹¹, 李继东², 李乐¹, 蒙国基¹, 于玉根¹

1. 深圳万乐药业有限公司深圳 518029;
2. 伯乐生命医学产品(上海)有限公司广州 510095

SNS:An Effective Strategy for Extending Ceramic Hydroxyapatite Column Life

DENG Yi-xi¹, LI Ji-dong², LI Le¹, MENG Guo-ji¹, YU Yu-gen¹

1. Shenzhen Main Luck Pharmaceuticals Inc., Shenzhen 518029, China;
2. Bio-Rad Laboratories(Shanghai) Ltd., Shenzhen 510095, China

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摘要：

陶瓷羟基磷灰石广泛用于各种生物分子的分离和纯化，但在层析应用上，纯化洗脱过程中填料表面氢原子的释放容易导致其性能和寿命降低。研究了一种可以延长CHT填料使用寿命的方法，通过在洗脱步骤前加入一种称为表面中和体系（SNS）的溶液，结合释放的氢原子，使得整个体系处于一个接近中性pH的状态。先筛选出有效调整pH的SNS条件，通过钙离子流失的监测证实该条件的有效性。再通过在WLB304单克隆抗体纯化工艺中加入SNS之后，对羟基磷灰石填料载量、pH和压力的变化、杂质的去除能力进行考察。结果表明，加入SNS可以有效减少钙离子流失，同时并不会影响CHT层析工艺的效果。

关键词： 陶瓷羟基磷灰石; SNS; 钙流失; 单克隆抗体

Abstract:

Ceramic Hydroxyapatite (CHT) has been widely used in the manufacturing of a wide variety of monoclonal antibodies (mAbs), vaccines, and other protein therapeutics. It is an effective tool for the removal of residual host cell proteins, nucleic acids, viruses, endotoxins, and product-related contaminants such as aggregates and degradation fragments. However, damage of the CHT media and increased column back pressure are often observed at production scale-up. A transient pH excursion during elution with increased salt concentration leads to Ca²⁺ depletion from the CHT column. In the present study,the effect of surface neutralization system (SNS) on the life cycles of CHT media is demonstrated. Following post-loading wash, a buffer with slightly elevated pH is applied onto the column so as to prevent significant pH drop during target protein elution. The level of Ca²⁺ depletion in response to pH and salt concentration of the SNS buffer have been investigated, and the optimal buffer composition for maintaining the pH of column effluent near neutral been identified. This newly developed protocol has been validated for the polish purification of mAb WLB304. The dynamic binding capacity of WLB304, impurity clearance efficiency, leached Ca²⁺ level, pH excursion and column back pressure during purification process were closely monitored. Data indicate that the implementation of SNS can significantly reduce Ca²⁺ depletion and effectively extend the life time of a CHT column, without compromising its chromatography performance.

Key words: SNS Antibody Calcium depletion CHT

收稿日期: 2016-07-25 出版日期: 2017-01-25

ZTFLH:

Q819

基金资助:

2014年度深圳市科技创新委员会技术攻关计划（深发改[2014]1841号）战略新兴产业发展专项资金资助项目（JSGG20141117163744687）

通讯作者: 蒙国基 E-mail: mengguoji@wanle.com.cn

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引用本文:

邓义熹, 李继东, 李乐, 蒙国基, 于玉根. 添加SNS能显著提高CHT填料使用寿命的研究[J]. 中国生物工程杂志, 2017, 37(1): 81-88.

DENG Yi-xi, LI Ji-dong, LI Le, MENG Guo-ji, YU Yu-gen. SNS:An Effective Strategy for Extending Ceramic Hydroxyapatite Column Life. China Biotechnology, 2017, 37(1): 81-88.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170112 或 https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I1/81

[1] Cummings L J, Ogawa T, Tunón P. CHT ceramic hydroxyapatite:new life for an old chromatographic technique.. http://www.bio-rad.com/webroot/web/pdf/lsr/literature/Bulletin_1927B.pdf.
[2] Nakamura S, Matsumoto T, Sasaki J, et al. Effect of calcium ion concentrations on osteogenic differentiation and hematopietic stem cell niche-related protein expressions in osteoblasts. Tissue Engineering, 2010,18(8):2467-2473.
[3] Yachelini P, Falk I, Eichmann K. Deviations in thymocyte development induced by divalent and monovalent ligation of the alpha/beta T cell receptor and by its cross-linking to CD8. J Immunol, 1990,145(5):1382-1389.
[4] Grun J B, et al. Viral removal/inactivation by purification of biopharmaceuticals. Biopharm, 1992,5:22-30.
[5] Juarez-Salinas H, et al. Separation of IgG idiotypes by high-performance hydroxylapatite chromatography. Methods Enzymol, 1986,121(58):615-622.
[6] Gagnon P, Paul N G, Jie H, et al. Platform purification of IgG with CHT ceramic hydroxyapatite.. www.ibclifesciences.com.
[7] Ng P K, Snyder M A. pH-based cation exchange chromatography in the capture and elution of monoclonal antibodies. J Sep Sci, 2012,35:29-35.
[8] Bankston T, Theresa E, Dattolo L,et al. pH transients in hydroxyapatite chromatography columns-Experimental evidence and phenomenological modeling. J of Chromatography A, 2010,1217:2123-2131.
[9] Belcher R, Close R A, West T S, et al. The complexometric titration of calcium in the presence of magnesium a critical study. Talanta, 1958,1:238-244.
[10] Kim J, Vipulanandan C. Effect of pH, sulfate and sodium on the EDTA titration of calcium. Cem Concr Res, 2003,33:621-627.
[11] Pabst T, Antos D, Carta G, et al. Protein separations with induced pH gradients using cation-exchange chromatographic columns containing weak acid groups. J of Chromatogr A, 2008,1181:83-94.

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