PURIFICATION OF PHASEOLUS VULGARIS LECTIN BY GEL FILTRATION CHROMATOGRAPHY
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It is well established that gel filtration chromatography is an effective technique for the purification of proteins by removing low-molecular-weight impurities. In this study, gel filtration chromatography was employed for the purification of lectin isolated from Phaseolus vulgaris L. seeds, with the aim of eliminating foreign low-molecular-weight and iodinated substances present in the crude extract. Lectin extraction was initially performed using a conventional saline buffer method. The purification process was carried out using a Bio-Rad BioLogic LP chromatography system equipped with a column packed with Sephadex G-75 gel. Tris-HCl buffer was used as the mobile phase, and the flow rate was maintained at 1.5 mL/min. The collected fractions were analyzed for protein content and lectin activity to identify lectin-rich fractions. The purified lectin was subsequently freeze-dried to obtain a solid form suitable for physicochemical characterization. Characterization included determination of protein and carbohydrate content, hemagglutination activity, and gel electrophoresis for molecular weight estimation. The results indicated successful purification of Phaseolus vulgaris lectin using gel filtration chromatography, yielding a biologically active lectin preparation with high purity. These findings confirm that Sephadex G-75 gel filtration is an efficient and reliable method for the purification of lectin from Phaseolus vulgaris seeds.
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