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Polyacrylamide/Alginate monolithic Cryogel-immobilized o-Phosphotyrosine (opt): purification of human IgG

Grant number: 16/10855-1
Support type:Regular Research Grants
Duration: November 01, 2016 - October 31, 2018
Field of knowledge:Engineering - Chemical Engineering
Principal Investigator:Sonia Maria Alves Bueno
Grantee:Sonia Maria Alves Bueno
Home Institution: Faculdade de Engenharia Química (FEQ). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil

Abstract

Antibodies, proteins with a very important use as human therapeutics, have also been employed in the analytical, diagnostic, biosensor, and proteomic areas. Their annual sales as therapeutics already exceed US$ 70 billion worldwide accounting for more than 6% of pharmaceutical industrial revenues. For these applications antibodies must have a high degree of purity. Since this required high purity are obtained with multistep, time consuming, high-cost, and low yield purification processes, there is a need for new purification technologies. One recent development in this field is monolith based chromatography in which the separation matrix is a single-unit material with a network of large interconnected pores. Due to this pore-structure the monotlith has a very low flow resistance and mass transfer is governed mainly by convection. The aim of this work is to develop a macroporous monolithic cryogel adsorbent based on polyacrylamide and alginate prepared under a constant freezing-temperature condition (cryo-polimerization) to be used to adsorb IgG from human serum. After its activation with epichorohydrin the affinity cryogel will be coupled to the pseudoaffinity ligand OPT since our group showed that his ligand (immobilized in agarose gel) showed high selectivity for IgG adsorption from human plasma (research financed by Fapesp). The starting materials used for the cryogel preparation are hydrophilic, cheap, and available. The physical properties, such as porosity, morphology, mechanical strength and hydraulic permeability of the cryogel will be characterized. The selectivity, the identification of Fc or Fab part of IgG adsorbed, and dynamic capacity of the adsorbent will also be. The equilibrium binding data will be determined to obtain thermodynamic parameters of adsorbents which are required for the development a large-scale process. (AU)

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