title: "Clickable" affinity ligands for effective separation of glycoproteins.
abstract: In this paper, we present a new modular approach to immobilize boronic acid ligands that can offer effective separation of glycoproteins. A new "clickable" boronic acid ligand was synthesized by introducing a terminal acetylene group into commercially available 3-aminophenyl boronic acid. The clickable ligand, 3-(prop-2-ynyloxycarbonylamino)phenylboronic acid (2) could be easily coupled to azide-functionalized hydrophilic Sepharose using Cu(I)-catalyzed 1,3-dipolar cycloaddition reaction under mild condition. Compared to other boronic acid affinity gels, the new affinity gel displayed superior effectiveness in separating model glycoproteins (ovalbumin and RNase B) from closely related bovine serum albumin and RNase A in the presence of crude Escherichia coli proteins. Because of the simplicity of the immobilization through "click chemistry", the new ligand 2 is expected to not only offer improved glycoprotein separation in other formats, but also act as a useful building block to develop new chemical sensors for analysis of other glycan compounds.
authors.affiliation: Department of Pure and Applied Biochemistry, Centre for Chemistry and Chemical Engineering, Lund University, Lund, Sweden.
authors.forename: Thummaruk
authors.initials: T
authors.lastname: Suksrichavalit
authors.forename: Keiichi
authors.initials: K
authors.lastname: Yoshimatsu
authors.forename: Virapong
authors.initials: V
authors.lastname: Prachayasittikul
authors.forename: Leif
authors.initials: L
authors.lastname: Bülow
authors.forename: Lei
authors.initials: L
authors.lastname: Ye
mesh_terms: D000818:Animals; D001386:Azides / Q000737:chemistry; D001897:Boronic Acids / Q000138:chemical synthesis / Q000737:chemistry*; D002417:Cattle; D002645:Chickens; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D029968:Escherichia coli Proteins / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D010047:Ovalbumin / Q000737:chemistry; D011485:Protein Binding; D012259:Ribonuclease, Pancreatic / Q000737:chemistry / Q000302:isolation & purification; D012260:Ribonucleases / Q000737:chemistry / Q000302:isolation & purification; D012685:Sepharose / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D001386:Azides; D001897:Boronic Acids; D029968:Escherichia coli Proteins; D006023:Glycoproteins; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; D012685:Sepharose; D012260:Ribonucleases; C029600:ribonuclease B; D012259:Ribonuclease, Pancreatic