Characterisation of a monoclonal antibody recognising specifically the HSP70 from Leishmania

Heat-shock protein 70 (HSP70) is ubiquitously distributed along the evoluti onary scale and has such an amino acid sequence conservation that it is con sidered the most evolutionarily conserved protein. In order to obtain immun ological tools specific against Leishmania infantum HSP70, hybridomas were established that secreted monoclonal antibodies (mAbs) against recombinant L. infantum HSP70. One of them, named mAb 2B8D2, specifically reacted with the Leishmania protein and did not recognise HSP70 from the related kinetop lastid Trypanosoma cruzi. The use of synthetic peptides allowed us to deter mine the B-cell epitope recognised by this mAb, an epitope located in the d ivergent C-terminal domain of the protein. Remarkably, the mAb possesses th e capacity to immunoprecipate HSP70 from promastigote extracts of L. infant um. The fact that human HSP70 is not recognised by this mAb assures the use fulness of this antibody for diagnostic purposes and studies involving Leis hmania infection of macrophages.