Lectins are proteins or glycoproteins, ubiquitous in nature, with specific and reversible carbohydrate binding and erythrocyte agglutination properties. Cratylia mollis is a  native forage from the State of Pernambuco, Northeast of Brazil, whose seeds contain lectin multiple molecular forms (Cramoll 1, 2, 3 and 4). The immobilization of proteins on insoluble supports produces affinity matrices. The aims of this work were immobilization of Cramoll 3 on Sepharose CL-4B and capacity evaluation of matrix (Cramoll 3-Sepharose) in binding glycoproteins. The isoform was obtained by previously established protocol. Cramoll 3 (2.8 mg) was immobilized on Sepharose CL-4B (2.5 mg). Glycoproteins (asialofetuin, 0.6 mg; azocasein, 1 mg; fetuin, 1 mg; ovoalbumin, 0.5 mg; thyroglobulin 0.5 mg) were applied to Cramoll 3-Sepharose columns (1 ml). Elutions were performed with 1 M NaCl and fractions were collected (1ml). The coupling of Cramoll 3 to Sepharose CL-4B was efficient (estimated 100 %). Columns did bind to asialofetuin and thyroglobulin (23%), as well as to azocasein (20%)  and ovoalbumin (10%); no binding was detected with fetuin. Cramoll 3-Sepharose was biospecifically selective when evaluated with different glycoproteins.