title: Purification of glycocalicin from human plasma.
abstract: Glycocalicin (GC) is a large extracellular proteolytic fragment of glycoprotein Ib, a membrane platelet component playing an essential role in the physiological processes of platelet adhesion and aggregation. GC contains the binding sites for thrombin and von Willebrand factor. GC circulates normally in vivo in significant concentrations and the plasma level of this protein reflects a complex function of factors including platelet count or platelet turnover. It can therefore serve as a good indicator for many diseases like hypoplastic thrombocytopenia and idiopathic thrombocytopenic purpura. For this reason, several purification assays have been previously described. In this work, we describe a novel analytical method for GC purification from human platelets based on preparative HPLC gel filtration followed by immuno-affinity chromatography on NHS activated column conjugated with specific antibody. Pure GC was obtained from tiny amount of starting material. Our protocol of GC purification is simple, fast and provides a pure end product.
authors.affiliation: Laboratoire de valorisation de la biomasse et production des protéines chez les eucaryotes, Centre de Biotechnologie de Sfax, Université de Sfax, Tunisia. Electronic address: hadjkacem_basma@yahoo.fr.
authors.forename: Basma
authors.initials: B
authors.lastname: HadjKacem
authors.affiliation: Laboratoire de valorisation de la biomasse et production des protéines chez les eucaryotes, Centre de Biotechnologie de Sfax, Université de Sfax, Tunisia.
authors.forename: Héla
authors.initials: H
authors.lastname: Mkaouar
authors.affiliation: Centre Régional de Transfusion Sanguine de Sfax- 99/UR/08-33, université de Sfax, Tunisia.
authors.forename: Ikram
authors.initials: I
authors.lastname: Ben Amor
authors.affiliation: Centre Régional de Transfusion Sanguine de Sfax- 99/UR/08-33, université de Sfax, Tunisia.
authors.forename: Jalel
authors.initials: J
authors.lastname: Gargouri
authors.affiliation: Laboratoire de valorisation de la biomasse et production des protéines chez les eucaryotes, Centre de Biotechnologie de Sfax, Université de Sfax, Tunisia.
authors.forename: Ali
authors.initials: A
authors.lastname: Gargouri
mesh_terms: D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000302:isolation & purification*
chemical_list: D019038:Platelet Glycoprotein GPIb-IX Complex; C023592:glycocalicin
keywords: Affinity chromatography; Blood purification; Glycocalicin purification; HPLC gel filtration; NHS column