Production and Standardization of Human Monoclonal Antibody Fragment (scFv) Expression in Tunair Shake Flask: Use of a Patented mAb - SERRUMAB

• Grant number: 24/17537-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: January 01, 2025
• End date: December 31, 2025
• Field of knowledge: Engineering - Biomedical Engineering - Bioengineering
• Principal Investigator: Manuela Berto Pucca
• Grantee: Julia Lopes de Oliveira
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

According to data from the Ministério da Saúde, in 2022 there were 183,738 scorpion accidents in Brazil, resulting in 92 deaths. The yellow scorpion (Tityus serrulatus), widely found in the country, is primarily responsible for severe cases. Although there is a specific treatment for these severe cases, it is still produced in animals (horses), which may lead to some disadvantages, such as anaphylaxis, serum sickness, and animal suffering. Therefore, it is still necessary to develop innovative strategies to neutralize this venom and produce a new generation of antivenoms, such as the use of monoclonal antibodies.This scientific initiation project aims to optimize the production of the human monoclonal antibody fragment SERRUMAB in E. coli HB2151, which recognizes the venom of Tityus serrulatus (yellow scorpion). The methodology includes comparative bacterial cultivation in conventional Erlenmeyer flasks and Tunair Shake flasks, which provide better oxygen transfer compared to the conventional ones, with cell growth monitored by optical density and dry weight measurements. After evaluating induction with lactose and IPTG, the cells will be processed to obtain the supernatant, which will then be purified. The cultivation performance will be evaluated through analyses of cell growth, purified antibody fragment concentration, and substrate consumption (lactose and glucose) by high-performance liquid chromatography (HPLC). The results will be analyzed after calculating kinetic parameters and confirming the maintenance of antibody recognition of the target (venom) by ELISA.The combination of process optimization with Tunair flasks and the application of engineering tools will provide a comprehensive understanding of the factors influencing SERRUMAB production, contributing to greater future scalability for the development of biotherapies targeting scorpion envenomation.