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筛选用于提高逆转录病毒和腺病毒载体稳定性的新型辅料。

Screening of novel excipients for improving the stability of retroviral and adenoviral vectors.

作者信息

Cruz Pedro E, Silva Ana Carina, Roldão António, Carmo Marlene, Carrondo Manuel J T, Alves Paula M

机构信息

IBET/ITQB, Apartado 12, P-2781-901 Oeiras, Portugal.

出版信息

Biotechnol Prog. 2006 Mar-Apr;22(2):568-76. doi: 10.1021/bp050294y.

Abstract

In the past decade there has been an increase in the application of viral vectors in the laboratory and clinical trials of human gene therapy, retroviral and adenoviral vectors among the most used. However, the limited stability of these vectors creates problems in the design of experiments, transport, and storage. Vectors stored at -80 degrees C must be quickly shipped on dry ice, which is somewhat cumbersome. Alternatively, viral vectors can be preserved in a lyophilized form. However, loss of viral activity during lyophilization can also be a serious problem. In this report we identify novel candidate formulations containing new compatible solutes, ectoin, hydroxyectoin, and firoin, that allow better stability of retroviral and adenoviral vectors during storage. For retroviral vectors, the maximum stabilization for long-term storage was achieved through lyophilization followed by storage at -20 degrees C using a formulation of Tris buffer pH 7.2 containing firoin (0.5 M), a half-life of 340 days being obtained. Adenoviral vectors storage at -80 degrees C in solution using Tris buffer pH 8.0 with firoin was the best method for long-term storage, with a half-life exceeding 1 year.

摘要

在过去十年中,病毒载体在人类基因治疗的实验室研究和临床试验中的应用有所增加,逆转录病毒载体和腺病毒载体是最常用的。然而,这些载体稳定性有限,给实验设计、运输和储存带来了问题。储存在-80℃的载体必须用干冰快速运输,这有些麻烦。或者,病毒载体可以冻干保存。然而,冻干过程中病毒活性的丧失也可能是一个严重问题。在本报告中,我们鉴定了含有新型相容性溶质(依克多因、羟基依克多因和丝胶蛋白)的新型候选配方,这些配方可使逆转录病毒和腺病毒载体在储存期间具有更好的稳定性。对于逆转录病毒载体,通过冻干,然后在-20℃下使用含有丝胶蛋白(0.5M)的pH7.2 Tris缓冲液配方储存,实现了长期储存的最大稳定性,半衰期为340天。腺病毒载体在含有丝胶蛋白的pH8.0 Tris缓冲液中于-80℃溶液中储存是长期储存的最佳方法,半衰期超过1年。

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