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通过基因免疫高效产生多克隆抗体。

High-level generation of polyclonal antibodies by genetic immunization.

作者信息

Chambers Ross S, Johnston Stephen Albert

机构信息

Center for Biomedical Inventions, Department of Internal Medicine, University of Texas-Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, Texas 75390-9185, USA.

出版信息

Nat Biotechnol. 2003 Sep;21(9):1088-92. doi: 10.1038/nbt858. Epub 2003 Aug 10.

Abstract

Antibodies are important tools for investigating the proteome, but current methods for producing them have become a rate-limiting step. A primary obstacle in most methods for generating antibodies or antibody-like molecules is the requirement for at least microgram quantities of purified protein. We have developed a technology for producing antibodies using genetic immunization. Genetic immunization-based antibody production offers several advantages, including high throughput and high specificity. Moreover, antibodies produced from genetically immunized animals are more likely to recognize the native protein. Here we show that a genetic immunization-based system can be used to efficiently raise useful antibodies to a wide range of antigens. We accomplished this by linking the antigen gene to various elements that enhance antigenicity and by codelivering plasmids encoding genetic adjuvants. Our system, which was tested by immunizing mice with >130 antigens, has shown a final success rate of 84%.

摘要

抗体是研究蛋白质组的重要工具,但目前生产抗体的方法已成为一个限速步骤。大多数产生抗体或抗体样分子的方法中的一个主要障碍是需要至少微克量的纯化蛋白。我们已经开发出一种利用基因免疫来生产抗体的技术。基于基因免疫的抗体生产具有几个优点,包括高通量和高特异性。此外,由基因免疫动物产生的抗体更有可能识别天然蛋白。在这里,我们表明基于基因免疫的系统可用于有效地产生针对多种抗原的有用抗体。我们通过将抗原基因与各种增强抗原性的元件连接并共同递送编码基因佐剂的质粒来实现这一点。我们的系统通过用超过130种抗原免疫小鼠进行了测试,最终成功率为84%。

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