Though　plant　lectins　have　been　widely　used　for　the　enrichment　of　glycoproteins/glycopeptides,　mammalian　lectins　have　rarely　been　explored　for　this　purpose.　Using　the　conserved　sequence　of　the　carbohydrate　recognition　domain　(CRD)　of　the　human　galectin-3　protein,　we　engineered　the　following　two　novel　human　galectins:Gal3C　(containing　one　CRD)　and　Tetra-Gal3C　(containing　four　tandem　CRDs).　Moreover,　we　designed　a　galectin　affinity　column　that　could　be　used　for　glycoprotein/glycopeptide　enrichment,　by　taking　advantage　of　the　ability　of　these　recombinant　galectins　to　become　immobilized　onto　streptavidin　agarose　beads.　SDS-PAGE,　western　blotting,　and　mass　spectrometry　were　used　to　characterize　the　biological　features　and　glycoprotein/glycopeptide　enrichment　abilities　of　both　these　recombinant　galectins.　Our　results　showed　that　both　Gal3C　and　Tetra-Gal3C　predominantly　enrich　glycoproteins/glycopeptides　and　that　this　enrichment　process　has　a　high　specificity　and　sensitivity.　Because　Tetra-Gal3C　contains　three　more　CRD　structural　domains,　it　exhibited　a　better　ability　to　achieve　enrichment　than　Gal3C.　Moreover,　the　immobilized　Gal3C　and　Tetra-Gal3C　were　successfully　used　for　the　enrichment　of　glycoproteins　from　HepG2　cells,　which　indicated　the　selective　glycoprotein/glycopeptide　enrichment　potential　of　recombinant　galectins　in　complex　samples.　©　2018　Chinese　Journal　of　Chromatography.