title: Role of tentacles and protein loading on pore accessibility and mass transfer in cation exchange materials for proteins.
abstract: In protein chromatography, the size of the protein determines which fraction of pores it can access within a resin and at which rate of diffusion. Moreover, in the presence of grafted polymers like in advanced materials, adsorbed proteins and electrolytes complicate the interaction pore-protein. In this study, we evaluated in a comparative way the behavior of Fractogel EMD SO3 (M) and (S), "tentacle"-type, strong cation exchangers, as well as a reference material without tentacles, all of which are commonly used for protein purification. ISEC experiments were carried out with a set of Dextran tracers of largely different molecular size covering the typical range of protein sizes. Experimental values of porosity (internal and external to the particles) as well as of pore diffusion coefficients have been measured at different NaCl concentrations and under protein loading. These results provide useful insights into the complex interplay among mentioned factors: first, the presence of tentacles induces size exclusion selectivity in the materials; second, the salt induces conformational changes of the tentacles, leading to porosities larger than expected in tentacle materials; third, protein adsorption mainly leads to a reduction of porosity due to pore space occupied by the protein and to a decrease of pore diffusion coefficient.
authors.affiliation: Merck KGaA, Frankfurter Str. 250, D-64293 Darmstadt, Germany.
authors.forename: Helen
authors.initials: H
authors.lastname: Thomas
authors.forename: Bertrand
authors.initials: B
authors.lastname: Coquebert de Neuville
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Storti
authors.forename: Massimo
authors.initials: M
authors.lastname: Morbidelli
authors.forename: Matthias
authors.initials: M
authors.lastname: Joehnck
authors.forename: Michael
authors.initials: M
authors.lastname: Schulte
mesh_terms: D000327:Adsorption; D000906:Antibodies / Q000737:chemistry*; D002412:Cations / Q000737:chemistry; D002852:Chromatography, Ion Exchange / Q000295:instrumentation* / Q000379:methods*; D003911:Dextrans / Q000737:chemistry; D004058:Diffusion; D008956:Models, Chemical; D008970:Molecular Weight; D011108:Polymers / Q000737:chemistry*; D016062:Porosity; D011205:Povidone / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry
chemical_list: D000906:Antibodies; D002412:Cations; D003911:Dextrans; D011108:Polymers; D011506:Proteins; D012965:Sodium Chloride; D011205:Povidone