Snake venoms are complex mixtures of proteins and peptides with different  biological activities, many of them very toxic. Several animals, including the opossum Didelphis marsupialis, are resistant to snake venoms due to the presence of neutralizing factors in their blood. An antihaemorrhagic protein named DM43 was isolated from opossum serum. It inhibits snake venom metalloproteinases through non covalent complex formation with these enzymes. In this study, we have used DM43 and proteomic techniques to explore snake venom subproteomes. Several venoms were chromatographed through an affinity column containing immobilized DM43. Bound fractions were analyzed either by SDS-PAGE and/or 2D-PAGE, followed by identification by MALDI-TOF/TOF MS. Following this methodology, we could classify venoms from Bothrops alternatus, B. asper, B. atrox, B. insularis, B. jararaca, B. jararacussu, B. moojeni, B. neuwiedi, Crotalus adamanteus, C. atrox, C. durissus terrificus, Lachesis muta muta and Naja naja atra according to their relative content of metalloproteinases (PI, PIII and/or their fragments). Venom fractions not bound to DM43 column were equally analyzed and were composed basically of serine proteinases, C-type lectins, L-amino acid oxidases, nerve growth factor, metalloproteinases and/or some unidentified spots. Studied venoms presented important proteic variability, with frequent detection of multiple forms of the same protein and several members of the same protein family. DM43 affinity chromatography associated with proteomic techniques showed to be useful tools to separate and identify proteins from snake venoms.