Precipitation as a purification method of monoclonal and polyclonal antibodies: determination of thermodynamic and process parameters

• Grant number: 20/12552-1
• Support Opportunities: Regular Research Grants
• Start date: March 01, 2022
• End date: February 29, 2024
• Field of knowledge: Engineering - Chemical Engineering - Industrial Operations and Equipment for Chemical Engineering
• Principal Investigator: Igor Tadeu Lazzarotto Bresolin
• Grantee: Igor Tadeu Lazzarotto Bresolin
• Host Institution: Instituto de Ciências Ambientais, Químicas e Farmacêuticas (ICAQF). Universidade Federal de São Paulo (UNIFESP). Campus Diadema. Diadema , SP, Brazil
• Associated researchers: Fabiana Perrechil Bonsanto ; Gisele Atsuko Medeiros Hirata ; Iara Rocha Antunes Pereira Bresolin ; Luís Antonio Peroni ; Rafael Mauricio Matricarde Falleiro
• Associated scholarship(s): 22/15751-0 - Evaluation of precipitation as a technique for the recovery and purification of anti-human IgM polyclonal antibodies from rabbit serum, BP.TT; 22/02807-8 - Production of anti-human IgG monoclonal antibodies and its purification by precipitation, BP.TT; 22/03417-9 - Evaluation of precipitation as a technique for the recovery and purification of anti-human IgM polyclonal antibodies from rabbit serum, BP.TT

Abstract

Antibodies are biomolecules that have become fundamental in several areas of study, such as clinical and diagnostics, due to their ability to recognize molecules present in a complex mixture. For the antibody purification step, precipitation stands out, which is based on the principle of reducing the solubility of a solute in a solvent, influenced by the addition of a precipitating agent, culminating in the formation of the precipitate in the medium under study. Thus, this project aims to study the use of precipitation as a technique to purify monoclonal (mAbs) and polyclonal (pAbs) anti-IgG and anti-IgM antibodies, respectively. The experiments will be carried out using polyethylene glycol and ZnCl2 as precipitating agents for these antibodies, determining their solubility curves and the most promising conditions for batch precipitation, as well as optimizing the resuspension step. After resuspension, it is sought an antibody with a purity higher than 90%, in a solution with a viscosity between 1 and 3 mPa s. Finally, the thermodynamic parameters binding constant (Ka), the reaction stoichiometry (n), and the variations of enthalpy (delta H), free energy (delta G) and entropy (delta S) will be determined by calorimetry, as well as the second virial coefficient (B22), which represents the degree of intermolecular interactions in diluted solutions. (AU)