High-level generation of polyclonal antibodies by genetic immunization.
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%.
Chambers
Ross S
RS
Center for Biomedical Inventions, Department of Internal Medicine, University of Texas-Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, Texas 75390-9185, USA. ross.chambers@utsouthwestern.edu
Johnston
Stephen Albert
SA
D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism*; D000917:Antibody Formation / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D007114:Immunization / Q000379:methods*; D051379:Mice; D014162:Transfection / Q000379:methods*
D000906:Antibodies