Functional recombinant human anti-HAV antibody expressed in milk of transgenic mice.
Functional recombinant human anti-HAV antibody expressed in milk of transgenic mice.
复制标题
在转基因小鼠的牛奶中表达的功能重组人抗HAV抗体。
DOI：
10.1007/s11248-008-9241-0
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发表时间：
2009-06
影响因子：
3
通讯作者：
Li N
中科院分区：
文献类型：
--
作者：
Zhang R;Rao M;Li C;Cao J;Meng Q;Zheng M;Wang M;Dai Y;Liang M;Li N
甲型肝炎病毒(HAV)是一种分布广泛的病原体，是世界范围内急性甲型肝炎的常见病因。甲型肝炎被动免疫在暴露后预防中发挥着极其重要的作用，临床应用往往需要大量抗体。作为一种替代体外生产重组蛋白的方法，在转基因动物的乳汁中表达单抗具有生产成本低、活性高等优点。在本论文中，通过联合显微注射编码中和抗体重链和轻链的两个盒，获得了8个转基因小鼠的建立系。Western blotting检测到表达的重链和轻链在乳腺中正确组装和修饰。抗体的高表达水平是在哺乳期实现的，并发现与整合转基因的拷贝数无关。经酶联免疫吸附试验和中和试验检测其结合特异性和中和活性，表明其能有效地中和甲型肝炎病毒JN株。因此，我们的研究结果表明，大规模高效生产转基因家畜乳汁中的抗甲型肝炎病毒单抗是可行的。
Hepatitis A virus (HAV) is a wide spread pathogenic agent and is the common cause of acute Hepatitis A worldwide. Passive immunization of HAV plays an extremely important role in post-exposure prophylaxis with clinical applications often requiring large amounts of antibody. As an alternative to the in vitro production of recombinant proteins, expression of monoclonal antibodies (mAbs) in the milk of transgenic animals is currently used being associated with low production costs and high activity. In this paper, eight founder lines of transgenic mice were generated by co-microinjection of the two cassettes encoding the heavy- and light-chains of a neutralizing anti-HAV antibody, respectively. The expressed heavy- and light-chains of the mAb were correctly assembled and modified in the mammary gland as detected by western blotting. High expression levels of the antibody were achieved during the lactation period and found to be independent of the copy numbers of integrated transgenes. The highest level was up to 32.2 mg/ml. The binding specificity and neutralizing activity of the expressed mAb were assayed by ELISA and neutralizing test, showing that it is capable to neutralize the JN strain of Hepatitis A virus efficiently. Therefore, our results suggest that a large-scale and efficient production of the anti-HAV mAb in the milk of transgenic farm animals would be feasible in the future.
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DOI：
10.1073/pnas.94.2.575
发表时间：
1997-01-21
影响因子：
11.1
作者：
Chung, JH;Bell, AC;Felsenfeld, G
通讯作者：
Felsenfeld, G
影响因子：
13.5
作者：
Martin, A;Lemon, SM
通讯作者：
Lemon, SM
影响因子：
9.4
作者：
MACGREGOR, A;KORNITSCHUK, M;GUST, ID
通讯作者：
GUST, ID
影响因子：
2.2
作者：
Newton, DL;Pollock, D;Rybak, SM
通讯作者：
Rybak, SM
影响因子：
3.7
作者：
Kim, SJ;Jang, MH;Hong, HJ
通讯作者：
Hong, HJ