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腺病毒载体在阴离子交换树脂上的色谱纯化

Chromatographic purification of adenoviral vectors on anion-exchange resins.

作者信息

Bo Huaben, Chen Jun, Liang Ting, Li Senhai, Shao Hongwei, Huang Shulin

机构信息

School of Bioscience and Biopharmaceutics, Guangdong Province Key Laboratory for Biotechnology Drug Candidates, Guangdong Pharmaceutical University, 510006 Guangzhou, Guangdong, PR China.

College of Pharmacy, Guangdong Pharmaceutical University, 510006 Guangzhou, Guangdong, PR China.

出版信息

Eur J Pharm Sci. 2015 Jan 25;67:119-125. doi: 10.1016/j.ejps.2014.11.004. Epub 2014 Nov 26.

Abstract

Anion-exchange chromatography is a useful and effective tool for adenoviral vectors purification. However, due to the different functional groups and matrices, both binding capacity and resolution of most AEC resins are different. In this study, four different AEC resins are compared by the binding capacity, resolution and recovery. Using Fractogel TMAE as an adsorbent to purify adenoviral vectors has obvious advantages over the other resins, namely (1) dynamic binding capacity is higher than other resins; (2) unprecedented sharpness (1,570,000±250,000) and symmetry of adenoviral vectors peak (1.67±0.06); (3) higher resolution with other contaminants (2.16±0.08); (4) no enzymatic treatment; (5) the recovery can reach 75%; (6) the purity is higher and the total virion to infectious particle ratios can reach 18.9. In the present work, we confirmed the possibility of purifying pharmaceutical-grade adenoviral vectors by AEC.

摘要

阴离子交换色谱法是用于腺病毒载体纯化的一种有用且有效的工具。然而,由于功能基团和基质不同,大多数阴离子交换色谱树脂的结合容量和分离度都有所不同。在本研究中,通过结合容量、分离度和回收率对四种不同的阴离子交换色谱树脂进行了比较。使用Fractogel TMAE作为吸附剂纯化腺病毒载体比其他树脂具有明显优势,即:(1)动态结合容量高于其他树脂;(2)腺病毒载体峰具有前所未有的尖锐度(1,570,000±250,000)和对称性(1.67±0.06);(3)与其他污染物的分离度更高(2.16±0.08);(4)无需酶处理;(5)回收率可达75%;(6)纯度更高,总病毒粒子与感染性粒子的比率可达18.9。在本研究中,我们证实了通过阴离子交换色谱法纯化药用级腺病毒载体的可能性。

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