Quantification of antibodies in affinity columns with proteins A and G: evaluation of the effects of viscosity on the performance of analytical methods

• Grant number: 22/07322-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: September 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: Amanda Calil Barcellos Leite
• Host Institution: Instituto de Ciências Ambientais, Químicas e Farmacêuticas (ICAQF). Universidade Federal de São Paulo (UNIFESP). Campus Diadema. Diadema , SP, Brazil

Abstract

Antibodies are biomolecules essential to several different surveys, studies, and clinical diagnoses due to their ability to recognize and link to specific molecules present in a complex mixture. In the process to obtain antibodies the phase of purification is a major step. The non-chromatographic technique of precipitation is highlighted among them. Precipitation is based on the principle of reducing the solubility of a solute in a solvent, influenced by the addition of a precipitating agent. Among those precipitating agents, it is remarkable polyethylene glycol (PEG), which has as a consequence an increase in viscosity, making the next steps extremely difficult. Thus, the goal of this project is the development, optimization, and validation of analytical methods in HPLC upon antibodies precipitation with polyethylene glycol evaluating the viscosity effects known as "viscous fingering". For this purpose, it will be used monoclonal (mAbs) and polyclonal (pAbs) antibodies using analytical columns POROS A and POROS G. For which one it will be assessed sturdiness parameters, level of specificity, precision, linearity, accuracy, detection, and measure thresholds.(AU)