Lectins are glycoproteins and/or oligomeric proteins found in a diverse assortment of organisms and have the extraordinary property of binding specifically, reversibly, and non-covalent to carbohydrates. Some lectins recognize sequences of two or more saccharides with specificity towards both inter-residue glycosidic linkages and anomeric configuration. For these reasons, lectins have proved to be useful reagents for the identification, isolation and characterization of the carbohydrate residues of glucoconjugates. Affinity chromatography based on the comercial resin sepharose CL-6B was used to isolate new Cl-b-type lectins from crude preparations of snake venoms (Bothrops jararaca, Bothrops jararacussu, Bothrops neuwiedi, Bothrops moojeni, Lachesis muta rhombeata). Most of the C-type lectins could be eluted with almost 100% recovery using the competitor isopropyl-b-D-thiogalactoside (IPTG) or through Ca²⁺ sequestration with EDTA. The lectins yield varied considerably among the diferent snake species, but Bothrops neuwied venom was a particularly rich source of lectins (2.7mg of lectin per mililiter of resin in saturating conditions). Cl-a-lectins from Crotalus durissus terrificus venom, from the jack fruit (jacalin) and the bread fruit seeds extract (frutalin) had no affinity, either with or without Ca²⁺ added, for Sepherose CL-6B, showing that the resin is specific for Cl-b-type lectins. The Sepharose CL-6B presents excelent proparties of gel filtration and as we have shown here, a new property to bind stereospecificity b-D-galactoside binding lectins-proteins. This new feature makes the method of Sepharose CL-6B-affinity chromatography an alternative for proteomic analysis and three-dimensional crystallographic studies since it is fast, reliable, and consequently amendable to easy scale-up. Other advantages of the technique are: high specificity for a certain carbohydrate structure compared to other affinity chromatography gels.