Study of culture conditions for production of scFv anti-Stx2 monoclonal antibody fragment using recombinant E. coli

• Grant number: 21/12282-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Effective date (Start): December 01, 2021
• Effective date (End): September 30, 2022
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Jose Geraldo da Cruz Pradella
• Grantee: Rafaela Vieira de Carvalho
• Host Institution: Instituto de Ciência e Tecnologia (ICT). Universidade Federal de São Paulo (UNIFESP). Campus São José dos Campos. São José dos Campos , SP, Brazil

Abstract

Shiga toxin-producing Escherichia coli (STEC) strains can cause hemorrhagic colitis and hemolytic uremic syndrome by producing Shiga cytotoxin (Stx), a toxin that inhibits protein synthesis by acting on rRNA, leading to cell death. As there is no available antidote, current therapy only involves treating symptoms. Monoclonal antibody fragments have become an important class of biopharmaceuticals currently available, presenting several advantages over the conventional antibody, such as its reduced size, ideal for large-scale production in microbial systems, robust processing technologies, and scalability for large-volume bioreactors. Recombinant scFv monoclonal antibody fragments against Stx toxins have been described as having neutralizing activity for diagnostic and therapeutic use against STEC. This project aims to study the obtainment of the scFv anti-Stx2 antibody fragment by varying the expression systems E. coli BL21 (DE3), BL21 (DE3) pays and Artic 10 recombinants and also through the optimization of the medium of culture and analysis of operational variables. (AU)