title: Gene-based vaccination and screening methods to develop monoclonal antibodies.
abstract: Gene-based in vivo electroporation has the potential to be used as a "protein-free" method to elicit immune responses and to generate monoclonal antibodies (mAb) against proteins/peptides in hosts. However, the method is very useful to raise mAbs against proteins and peptides and not for carbohydrates, lipids, or haptens. Nevertheless, making mAb using this potent method faces a few challenges: the parameters of the electroporation needs further standardized, the final boost still needs protein antigens, and the primary screening of the clones requires purified protein. We present methods to overcome these challenges by an optimized electroporation framework and a method to use transiently transfected cells for the final boost, as well as for screening of the resulting clones via the use of an "In-Cell Western" method.
authors: Department of Microbiology and Immunology, University of Miami, Miami, FL, USA.
authors: M Pirouz
authors: MP
authors: Daftarian
authors: Aram
authors: A
authors: Vosoughi
authors: Vance
authors: V
authors: Lemmon
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D019556:COS Cells; D002522:Chlorocebus aethiops; D018274:Electroporation; D005260:Female; D006825:Hybridomas / Q000166:cytology; D051379:Mice; D008807:Mice, Inbred BALB C; D016707:Tissue Fixation; D014162:Transfection; D014611:Vaccination / Q000379:methods*; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D019444:Vaccines, DNA