title: One-step coating of silica capillaries for selective protein retention by Cu(II)-IDA IMAC.
abstract: A simple protocol to obtain Cu(II)-IDA (iminodiacetic acid)-modified capillaries was developed for immobilized metal ion affinity chromatography (IMAC). It consisted in the synthesis of IDA-silane used for a one-step coating of fused silica capillaries. The approach prevented the hydrolysis of silica potentially induced by two step coatings (γ-GPTMS, then IDA) employed in the conventional method of bonding iminodiacetic acid. The IDA content was quantified using a model relating the electroosmotic flow generated in IDA-modified capillaries to the charges induced by IDA species. The retention behavior of holotransferrin and bovine serum albumin on these IMAC columns was then investigated. Holotransferrin revealed a high affinity for Cu(II)-supports through a specific interaction with Cu(II) ions whereas albumin did not show any retention. The use of such columns for sample pretreatment before an HPLC analysis was proved to be successful.
authors.affiliation: Laboratoire de Chimie Analytique et Sciences Séparatives IPHC, Département Sciences Analytiques, Université de Strasbourg, CNRS, 25 rue Becquerel, F-67087 Strasbourg Cedex 2, France. zeinamehyou@hotmail.com
authors.forename: Zeina
authors.initials: Z
authors.lastname: Mehyou
authors.forename: Ryszard
authors.initials: R
authors.lastname: Lobinski
authors.forename: Agnès
authors.initials: A
authors.lastname: Hagège
mesh_terms: D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000295:instrumentation*; D003300:Copper / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000295:instrumentation; D006801:Humans; D007098:Imino Acids / Q000138:chemical synthesis / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000302:isolation & purification*; D012821:Silanes / Q000138:chemical synthesis / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry*; D014168:Transferrin / Q000302:isolation & purification*
chemical_list: D007098:Imino Acids; D012821:Silanes; D014168:Transferrin; C110714:holotransferrin; D012710:Serum Albumin, Bovine; D012822:Silicon Dioxide; D003300:Copper; C008109:iminodiacetic acid