GENERATION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES SPECIFIC FORMEMBERS OF THE MAMMALIAN 70-KDA HEAT-SHOCK PROTEIN FAMILY

The 70-kDa heat shock proteins (hsp70) are a highly conserved, abundan t, and ubiquitous family of proteins expressed by all organisms from b acteria to humans, It is well established that hsp70 family members fu nction as molecular chaperones and aid in the intracellular folding of newly synthesized or denatured proteins, Current evidence suggests an emerging role for hsp70 family members in immune responses and in cli nically important responses to stress and tissue damage, Here we repor t the generation and characterization of several MAbs to hsp70 family members, Immune responses to this highly conserved family were induced in mice by immunization with synthetic peptides that contain regions of the mouse mitochondrial hsp7O coupled to a potent helper T cell epi tope derived from tetanus toroid, The resulting MAbs include ones spec ific for the human and mouse mitochondrial hsp70 and others that show cross-reactivity among the family members and recognize the mitochondr ial hsp70, the endoplasmic reticulum resident hsp70, Bip/grp78 the con stitutively expressed cytosolic hsp70, hsc70, and the heat-induced mem ber, hsp70, Significantly, these MAbs are effective in Western blottin g, in immunoprecipitation, and in immunofluorescence, and thus should find applications in the purification and detection of members of this important family.