An improved and robust DNA immunization method to develop antibodies against extracellular loops of multi-transmembrane proteins.

ABSTRACT: Multi-transmembrane proteins are especially difficult targets for antibody generation largely due to the challenge of producing a protein that maintains its native conformation in the absence of a stabilizing membrane. Here, we describe an immunization strategy that successfully resulted in the identification of monoclonal antibodies that bind specifically to extracellular epitopes of a 12 transmembrane protein, multi-drug resistant protein 4 (MRP4). These monoclonal antibodies were developed following hydrodynamic tail vein immunization with a cytomegalovirus (CMV) promoter-based plasmid expressing MRP4 cDNA and were characterized by flow cytometry. As expected, the use of the immune modulators fetal liver tyrosine kinase 3 ligand (Flt3L) and granulocyte-macrophage colony-stimulating fact

SUBMITTER: Hazen M 

PROVIDER: S-EPMC3929459 | biostudies-literature | 2014 Jan-Feb

REPOSITORIES: biostudies-literature