Three different lipases from the extract crude of Staphylococcus warneri have been purified by specific lipase–lipase interactions using different lipases (TLL, RML, PFL, BTL2) covalently attached to a solid support as adsorption matrix. BTL2 immobilized on glyoxyl-DTT adsorbed selectivity only a 30kDa lipase from the crude, which was desorbed by adding 0.1% triton X-100. Using glyoxyl-PFL as matrix, two new lipases (28 and 40kDa) were adsorbed, and completely pure 40kDa lipase was obtained after desorption using 0.01% triton, whereas 28kDa lipase was desorbed after the incubation of the lipase matrix with 3% detergent. When using other matrixes as glyoxyl-TLL or glyoxyl-RML, different lipases were adsorbed. This methodology could be a very efficient and useful method to purify several lipases from crude extracts from different sources.