title: Preparation and testing of stationary phases and modified capillaries for affinity chromatography and affinity capillary electrophoresis of pepsin.
abstract: Three stationary phases have been prepared for affinity liquid chromatography isolation and separation of porcine and human pepsin. The phases contain 3,5-diiodo-L-tyrosine (DIT) bound to the supports HEMA BIO VS, HEMA BIO E and EPOXY TOYOPEARL. These phases have been tested on a model sample of porcine pepsin A and applied to human pepsin. Fractions have been collected and the chymase activity determined in selected analyses. For affinity CE, capillaries have been prepared by modifying the wall with 3-aminopropyltriethoxysilane, followed either by direct binding of DIT, or by binding L-tyrosine that was subsequently iodated. The dissociation constant K(d) has been determined for the pepsin-DIT complex from the changes in the electrophoretic mobilities.
authors.affiliation: Department of Analytical Chemistry, Charles University, Prague, Hlavova 2030/8, Praha 2, 12843 Czech Republic. tereza.varilova@seznam.cz
authors.forename: Tereza
authors.initials: T
authors.lastname: Varilová
authors.forename: Alice
authors.initials: A
authors.lastname: Vránková
authors.forename: Vera
authors.initials: V
authors.lastname: Pacáková
authors.forename: Marie
authors.initials: M
authors.lastname: Tichá
authors.forename: Karel
authors.initials: K
authors.lastname: Stulík
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods; D004105:Diiodotyrosine / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D010434:Pepsin A / Q000302:isolation & purification*; D012822:Silicon Dioxide / Q000737:chemistry; D013552:Swine
chemical_list: D004105:Diiodotyrosine; D012822:Silicon Dioxide; D010434:Pepsin A