Effect of the spacer arm on non-specific binding in membrane affinity chromatography
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Research Letter
Effect of the spacer arm on non-specific binding in membrane affinity chromatography Eleonora Lalli, Giulio C. Sarti, and Cristiana Boi, Dipartimento di Ingegneria Civile, Chimica, Ambientale e dei Materiali, DICAM, Alma Mater Studiorum-Università di Bologna, via Terracini 28, 40131 Bologna, Italy Address all correspondence to Cristiana Boi at [email protected] (Received 16 September 2017; accepted 3 January 2018)
Abstract The preparation, screening, and characterization of affinity membranes require a deep knowledge of the behavior of all components of the affinity material. Several studies report the effect of different spacers in combination with the ligand molecule, but the effect of the spacer arm “per se” is generally disregarded. The effect of the spacer 1,2-diaminoethane on non-specific protein adsorption was recently investigated and the results were compared with the ones obtained with A2P affinity membranes. The results show that this spacer has indeed an important effect and that similar specific studies need to be performed for every spacer molecule.
Introduction Affinity technology is a fundamental method that allows selective and efficient chromatographic purification of bioproducts by exploiting the specific and reversible interaction between a ligand, immobilized on a solid support, and the target molecule. To this aim, any other non-covalent interactions such as hydrogen bonding and hydrophobic interactions that may arise between any molecule in solution and any part of the derivatized affinity support should be minimized. Due to the high cost of affinity materials, the industrial application of this technology is considered feasible only for the purification of high added value products such as biotherapeutics. Among those, monoclonal antibodies (mAbs) are the class of therapeutics that arouse more interest due to their importance in the cure of illnesses such as cancer, autoimmune diseases, rheumatoid arthritis, and several other diseases.[1] The development of a platform for antibodies purification is due to the exploitation of the interaction between Protein A and the constant fragment, the so-called Fc moiety, of the antibody.[2] This specific interaction allows the purification of a whole class of mAbs with the same process, regardless of their individual antigenic activity.[3,4] Indeed, the high cost of this process step explains the enormous research effort in this area to find economical alternatives to Protein A affinity chromatography.[5–9] Within this framework, the development of affinity membranes with synthetic ligands, that mimic the interaction between Protein A and the Fc moiety of immunoglobulins is being investigated.[10,11] Although the major attention is commonly devoted to the effects of different ligands, the purification performance of an affinity membrane depends on several factors, somehow related one to the other, i.e. the
membrane material and structure, the affinity ligand, the spacer arm, and the immobilization protocol applied. The desired characteristics of the me
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