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High-level generation of polyclonal antibodies by genetic immunization

Nature Biotechnology volume 21, pages 1088–1092 (2003)Cite this article

Abstract

Antibodies are important tools for investigating the proteome, but current methods for producing them have become a rate-limiting step1. 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 immunization2. Genetic immunization–based antibody production offers several advantages, including high throughput3 and high specificity. Moreover, antibodies produced from genetically immunized animals are more likely to recognize the native protein2. 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%.

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Figure 1: Design of genetic immunization vector.
Figure 2: Antibody responses of mice immunized with pBQAP10-AAT.
Figure 3: Western blot analysis of natural extracts.

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Acknowledgements

We thank Ryan Anholt, Anwarul Ferdous, Kyle Meinert, Joey Nguyen, Vickie Seward, Rebecca Solis and Greg Urquhart for technical assistance, Mike McGuire, Bao-Xi Qu and Irene Rombel for helpful discussions and Rick Lyons for supplying the Mtb extracts. This work was supported by grants from the Programs for Genomic Applications (PGA) from the US National Heart, Lung, and Blood Institute to S.A.J. at the University of Texas Southwestern (U01HL66880), and from the Countermeasures Program from the University of Texas Austin.

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

    Ross S Chambers & Stephen Albert Johnston

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  1. Ross S Chambers
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  2. Stephen Albert Johnston
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Correspondence to Ross S Chambers.

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Chambers, R., Johnston, S. High-level generation of polyclonal antibodies by genetic immunization. Nat Biotechnol 21, 1088–1092 (2003). https://doi.org/10.1038/nbt858

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