title: A comprehensive evaluation of mixed mode interactions of HEA and PPA HyperCel™ chromatographic media.
abstract: Mixed mode or multimodal chromatography has been developed for rational use of multiple interactions in a controlled manner, in contrast to non-specific interactions. Indeed, as the term "mixed mode" suggests, these resins allow different types of interactions within a single chromatographic medium. In this paper, HEA HyperCel™, PPA HyperCel™ mixed-mode chromatographic media have been studied. These mixed-mode sorbents typically involve hydrophobic pseudo-affinity interactions for binding and essentially ionic interactions (charge repulsion) for elution. We identified and characterized these different interactions in chromatographic experiments by exploiting specific properties of proteins using protein standards and complex mixtures. We highlighted the major intervention of at least two types of interactions in these media: hydrophobic and electrostatic interactions. We observed the behaviour of these resins at different pH, ionic strength, with different salts and buffers types and in the presence of different organic compounds.
authors.affiliation: IPB, Biotechnologie des Protéines Recombinantes à Visée Santé, EA 4135, F-33000 Bordeaux, France; Univ Bordeaux, Biotechnologie des Protéines Recombinantes à Visée Santé, EA 4135, F-33000 Bordeaux, France; Pall Life Sciences, 48 Avenue des Genottes, 95800 Cergy Saint-Christophe, France.
authors.forename: J
authors.initials: J
authors.lastname: Pezzini
authors.affiliation: IPB, Biotechnologie des Protéines Recombinantes à Visée Santé, EA 4135, F-33000 Bordeaux, France; Univ Bordeaux, Biotechnologie des Protéines Recombinantes à Visée Santé, EA 4135, F-33000 Bordeaux, France.
authors.forename: C
authors.initials: C
authors.lastname: Cabanne
authors.affiliation: Pall Life Sciences, 50 Bearfoot Road, Northborough, MA 01532, USA.
authors.forename: R
authors.initials: R
authors.lastname: Gantier
authors.affiliation: CBST, VIT University, Vellore, Tamil Nadu, India.
authors.forename: V N
authors.initials: VN
authors.lastname: Janakiraman
authors.affiliation: IPB, Biotechnologie des Protéines Recombinantes à Visée Santé, EA 4135, F-33000 Bordeaux, France; Univ Bordeaux, Biotechnologie des Protéines Recombinantes à Visée Santé, EA 4135, F-33000 Bordeaux, France. Electronic address: xavier.santarelli@ipb.fr.
authors.forename: X
authors.initials: X
authors.lastname: Santarelli
mesh_terms: D002845:Chromatography / Q000379:methods*; D057927:Hydrophobic and Hydrophilic Interactions; D011506:Proteins / Q000737:chemistry
chemical_list: D011506:Proteins
keywords: HEA HyperCel™; Mixed mode chromatography; PPA HyperCel™; Proteins; Purification