Overcoming the poor immunogenicity of a protein by DNA immunization as a fusion construct.
The production of antibodies against poorly immunogenic proteins is problematic. Often there is a failure to generate such antibodies. Furthermore, antibodies against other specificities are frequently induced. We describe a simple approach, analogous to conjugation to a protein carrier, whereby immunization with naked DNA was used to raise antibody to a highly homologous and poorly immunogenic allotypic protein. Deoxyribonucleic acid encoding the protein of interest was fused to DNA encoding the Fc region of a foreign Ig, resulting in increased immunogenicity. The potential applications of this approach include the production of antisera and mAb to allotypic variants, mutant proteins, and proteins that are highly conserved between species.
Sutherland
R M
RM
The Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia.
McKenzie
B S
BS
Corbett
A J
AJ
Brady
J L
JL
Lew
A M
AM
D000818:Animals; D000917:Antibody Formation; D000941:Antigens / Q000276:immunology* / Q000378:metabolism; D019556:COS Cells; D002469:Cell Separation; D005434:Flow Cytometry; D006801:Humans; D007114:Immunization / Q000379:methods*; D007126:Immunoglobulin Allotypes / Q000276:immunology; D017493:Leukocyte Common Antigens / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008856:Microscopy, Fluorescence; D010957:Plasmids; D013950:Thymus Gland / Q000166:cytology; D014162:Transfection; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology*
D000941:Antigens; D007126:Immunoglobulin Allotypes; D019444:Vaccines, DNA; D017493:Leukocyte Common Antigens