Production of scFv-Fc fusion protein using genetically manipulated quails
Production of scFv-Fc fusion protein using genetically manipulated quails
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DOI：
10.1263/jbb.102.297
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发表时间：
2006-10-01
影响因子：
2.8
通讯作者：
Iijima, Shinji
中科院分区：
文献类型：
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作者：
Kawabe, Yoshinori;Kamihira, Masamichi;Iijima, Shinji
已经提出使用转基因禽类物种作为转基因生物反应器用于生产重组蛋白。近年来，虽然已经报道了用于产生转基因鸡的各种方法，但是很少达到以商业上可行的水平表达有用的蛋白质。在这项研究中，我们将浓缩的逆转录病毒载体注射到鹌鹑胚胎中，以产生产生重组蛋白的基因操纵鹌鹑。我们发现，在整个身体中的转基因表达在一个高的水平，观察到病毒注射到心脏的胚胎发育48小时孵育后。为了实际生产有用的蛋白质，将编码在P-肌动蛋白启动子控制下的抗朊病毒scFv-Fc基因的逆转录病毒载体注射到鹌鹑胚胎中。稳定孵化的鹌鹑在其血清和蛋白色中以高水平产生scFv-Fc。scFv-Fc的产生在整个育种期间保持。从白色蛋白中纯化的scFv-Fc保留了抗原结合活性。该系统显示了转基因鹌鹑用于商业化生产重组蛋白的潜力。
The use of transgenic avian species as a transgenic bioreactor for the production of recombinant proteins has been proposed. In recent years, although various procedures for generating transgenic chickens have been reported, the expression of a useful protein at a commercially feasible level has rarely been attained. In this study, we injected a concentrated retroviral vector into quail embryos to generate genetically manipulated quails that produce recombinant proteins. We found that transgene expression in the whole body at a high level was observed for viral injection into the heart of the developing embryos after a 48-h incubation. For the practical production of a useful protein, a retroviral vector encoding an anti-prion scFv-Fc gene under the control of the P-actin promoter was injected into quail embryos. The quails that hatched stably produced scFv-Fc at a high level in their serum and egg white. The production of scFv-Fc was maintained throughout the breeding period. scFv-Fc purified from the egg white retained the antigen-binding activity. This system exhibited the potential of transgenic quails for the commercial production of recombinant proteins.