title: Adsorption and separation of proteins by a synthetic hydrotalcite.
abstract: In this study, the potential use of a synthetic Mg/Al hydrotalcite (layered double hydroxide) as a novel chromatography material for protein purification was investigated. The hydrotalcite is present in its carbonate form and is characterized by an Al/Mg-ratio of 1.85. Zetapotential measurements confirm a positive surface potential up to pH 10 suggesting applicability as anion exchanger. The binding of model proteins covering a broad range of isoelectric points and molecular weights was performed at different pH-values under batch conditions to evaluate the binding behaviour of the hydrotalcite. Furthermore, static binding capacities were exemplarily determined for hemoglobin and human serum albumin. Additionally, the adsorption and elution of hemoglobin was studied under dynamic conditions. The binding behaviour of the hydrotalcite was compared to commercially available anion exchangers and was found to be a function of pH, depending on the model protein. Variant adsorption behaviour is explained by further interactions like hydrogen bonds and by an unequal charge distribution over the protein surfaces. The hydrotalcite reveals high adsorption capacities under static (260 mg/g) as well as under dynamic conditions (88 mg/g at 34 cm/h; 61 mg/g at 340 cm/h). With appropriate buffers like 500 mM carbonate (pH 10) the adsorbed proteins can be nearly completely desorbed making regeneration possible. Due to the binding and elution properties it is concluded, that the hydrotalcite can serve anion exchange material for chromatographic protein separations.
authors.affiliation: Institut für Biotechnologie, FG Bioverfahrenstechnik, Technische Universität Berlin, Berlin, Germany. kathrin.ralla@web.de
authors.forename: Kathrin
authors.initials: K
authors.lastname: Ralla
authors.forename: Ulrich
authors.initials: U
authors.lastname: Sohling
authors.forename: Kirstin
authors.initials: K
authors.lastname: Suck
authors.forename: Friederike
authors.initials: F
authors.lastname: Sander
authors.forename: Cornelia
authors.initials: C
authors.lastname: Kasper
authors.forename: Friedrich
authors.initials: F
authors.lastname: Ruf
authors.forename: Thomas
authors.initials: T
authors.lastname: Scheper
mesh_terms: D000327:Adsorption; D000536:Aluminum Hydroxide / Q000737:chemistry* / Q000378:metabolism; D000838:Anions / Q000737:chemistry* / Q000378:metabolism; D002852:Chromatography, Ion Exchange / Q000379:methods*; D017743:Equipment Reuse; D006454:Hemoglobins / Q000378:metabolism*; D006801:Humans; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008276:Magnesium Hydroxide / Q000737:chemistry* / Q000378:metabolism; D008970:Molecular Weight; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*
chemical_list: D000838:Anions; D006454:Hemoglobins; D012709:Serum Albumin; C010467:hydrotalcite; D000536:Aluminum Hydroxide; D008276:Magnesium Hydroxide