TY - JOUR
T1 - High-level generation of polyclonal antibodies by genetic immunization
AU - Chambers, Ross S.
AU - Johnston, Stephen Albert
N1 - Funding Information:
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.
PY - 2003/9
Y1 - 2003/9
N2 - 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%.
AB - 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%.
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U2 - 10.1038/nbt858
DO - 10.1038/nbt858
M3 - Letter
C2 - 12910245
AN - SCOPUS:0041859398
SN - 1087-0156
VL - 21
SP - 1088
EP - 1092
JO - Nature biotechnology
JF - Nature biotechnology
IS - 9
ER -