Aqueous two-phase micellar systems as a new methodology to recover recombinant antibody fragment from Pichia pastoris fermented broth

PELLEGRINI MALPIEDI, LUCIANA; DIAZ, CÉSAR A; NERLI, BIBIANA; PESSOA JR, ADALBERTO

Portland

Simposio; 35th Symposium on Biotechnology for Fuels and Chemicals.; 2013

Recombinant antibody fragments (scFv) are becoming one of the most popular tools in human therapies since they possess several advantageous features as easy manipulation, fast elimination of their inmunocomplex and the possibility of being produced in simple expression systems like bacteria and yeast. The objective of this work is to evaluate the application of aqueous two-phase micellar systems (ATPMS) of triton X-114 to recover an anti-LDL scFv produced extracellularly by Pichia pastoris. Cloud points were determined to construct the coexistence curves in buffered solutions of pH 7.4, with or without NaCl, Cibacrom Blue 3FGA (CB) and FabsorbentTM F1P HF (HF) ligand, and yeast fermented broth (60% of the total system mass). scFv partition was studied following a 23-full factorial design in which NaCl, CB and HF concentrations were selected as independent variables, and scFv purification factor (PF) and percentage yield (Y%) as responses. Triton concentration (2% w/w) and temperature (17°C) were maintained constant.