Purification of Lectin in Hemolymph of Sarcophaga peregrina Larvae by Immuno affinity Chromatography

A method of immuno affinity chromatography for insect lectin purification was described.Antibody against lectin in the hemolymoph of Sarcophaga peregrina larvae was raised by immunizing male rabbits and the complex of lectin binding with to rabbit blood red cells was used as antigen.Then the antibody used as immuno affinity ligand was coupled onto the matrix Sepharose 4B.The lectin with M r 73 kD in the hemolymph of S.peregrina larvae was purified through immuno affinity chromatography.The purified lectin was shown the same molecular weight as that purified from general affinity chromatography by using two glycoproteins as affinity ligands,which could inhibit the activity of this lectin.Comparison of these three methods indicated that the way of this immuno affinity chromatography did work well in purification of lectins.Immuno affinity chromatography would be an effective and an alternative way for insect lectin purification,especially when the specific binding saccharides of the lectins were unknown or atypical.