expression, purification and characterization of human recombinant galectin 3 in pichia pastoris

background: over the past century, the areas of genomics, proteomics and lipids have captured the attention of investigators worldwide. carbohydrates, have recently received increased attention through the expanding field of glycobiology; probably because they are very complex and not encoded in the genome. objectives: the purpose of this study was to express and purify recombinant human galectin 3via the pichiapastoris expression system. materials and methods:cdna of human galectin 3 gene was amplified with specific primers and cloned into a pcdna3.1 vector with his-tag for easier purification using ni2andchromatography. furthermore, galectin 3was purified to homogeneity and confirmed using sds-page and western blotting. results:the protein band corresponding to 29 kda was excised from the gels, digested with trypsin and processed for mass spectrometric analysis by matrix assisted laser desorption/ionization- time of flight mass spectroscopy (maldi-tof ms), using a reflex iii instrument. conclusions:tryptic digest analysis clearly revealed that the purified protein was indeed galectin 3. similarly, the biological activity of recombinant galectin 3 was confirmed using the hemagglutination inhibition assay.