The global prevalence of individuals with lactose intolerance and galactosemia has created a new market for lactose-free food products. In this scenario, the use of systems containing molecules with the ability to capture lactose is very promising. Therefore, a lectin from papaya seeds with the ability to bind lactose was extracted, purified and characterized. This lectin was immobilized in polyaniline and used as a bioaffinity column to remove this disaccharide.