Extraction of monoclonal antibody fragments type scFv by aqueous two-phase micellar systems

• Grant number: 11/20521-0
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: April 01, 2012
• End date: March 31, 2014
• Field of knowledge: Engineering - Chemical Engineering - Chemical Process Industries
• Principal Investigator: Adalberto Pessoa Junior
• Grantee: Luciana Pellegrini Malpiedi
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Due to a high percentage (70-80% correspond to the separation and purification steps) of the costs to obtain biotechnological products, it is necessary to optimize some methodologies to ensure a good cost/benefit relationship. Nowadays, the micellar aqueous two-phases systems are considered as an appropriated bio-separating technology because their properties are suitable to maintain the macromolecule's integrity and also allow regulating the experimental conditions enabling a selective extraction. In this project, the liquid-liquid extraction by micellar aqueous two-phase systems, composed by Triton X-114/ buffer McIlvaine, will be studied as a method to separate and to purify scFv antibodies from Pichia pastoris cultures. The effect of the system's components on the antibody integrity will be studied as well as the effect of different variables as pH, temperature, surfactant concentration, salts concentration and the presence of ligand molecules over the partition and purification of the scFv antibody. Finally, the best experimental conditions will be selective to apply a 1000-times scaling in order to test their potential for industrial application. (AU)

Scientific publications

(5)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

MALPIEDI, LUCIANA P.; NERLI, BIBIANA B.; ABDALLA, DULCINEIA S. P.; PESSOA, JR., ADALBERTO. Assessment of the effect of triton X-114 on the physicochemical properties of an antibody fragment. BIOTECHNOLOGY PROGRESS, v. 30, n. 3, p. 554-561, MAY-JUN 2014. (11/20521-0)

LARIO, LUCIANA DANIELA; PELLEGRINI MALPIEDI, LUCIANA; BRANDAO PEREIRA, JORGE FERNANDO; SETTE, LARA DURAES; PESSOA-JUNIOR, ADALBERTO. Liquid-liquid extraction of protease from cold-adapted yeast Rhodotorula mucilaginosa L7 using biocompatible and biodegradable aqueous two-phase systems. SEPARATION SCIENCE AND TECHNOLOGY, v. 51, n. 1, p. 57-67, JAN 2 2016. (13/19486-0, 11/20521-0, 12/23726-4)

MALPIEDI, LUCIANA P.; NERLI, BIBIANA B.; ABDALA, DULCINEIA S. P.; PESSOA-FILHO, PEDRO DE ALCANTARA; PESSOA, JR., ADALBERTO. Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purification. Separation and Purification Technology, v. 132, p. 295-301, AUG 20 2014. (11/20521-0)

VICENTE, FILIPA A.; MALPIEDI, LUCIANA P.; E SILVA, FRANCISCA A.; PESSOA, JR., ADALBERTO; COUTINHO, JOAO A. P.; VENTURA, SONIA P. M.. Design of novel aqueous micellar two-phase systems using ionic liquids as co-surfactants for the selective extraction of (bio)molecules. Separation and Purification Technology, v. 135, p. 259-267, OCT 15 2014. (12/12022-6, 11/20521-0)

MALPIEDI, LUCIANA P.; NERLI, BIBIANA B.; TAQUEDA, MARIA E. S.; ABDALLA, DULCINEIA S. P.; PESSOA, JR., ADALBERTO. Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systems. Protein Expression and Purification, v. 111, p. 53-60, JUL 2015. (11/20521-0)