Expression and Characterization of an scFv Antibody Fragment Inhibitor of Human Tissue Kallikrein 7

• Grant number: 25/04504-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: June 01, 2025
• End date: May 31, 2026
• Field of knowledge: Biological Sciences - Biochemistry - Enzymology
• Principal Investigator: Vitor Marcelo Silveira Bueno Brandão de Oliveira
• Grantee: Samara Evangelista da Silva
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

Atopic Dermatitis (AD) is a disease that affects approximately 7.3% of the Brazilian population aged 6 to 7 years and has a prevalence of about 1% to 3% of the adult global population. AD is characterized by three main symptoms: excessive skin scaling, pruritus, and erythema (redness). It is a chronic disease that tends to diminish over the individual's lifetime. However, in some cases, the disease may persist throughout the patient's life, requiring constant treatment during flare-ups. Studies indicate that the exacerbated activity of serine proteases from the kallikrein family (hKLKs), especially hKLK7 and hKLK5, contributes to the development and worsening of AD. Therefore, drugs that control the activity of these enzymes are of great interest for the treatment of this disease. In healthy individuals, hKLK7, along with other proteases from the kallikrein family, plays a role in the natural desquamation of the stratum corneum. However, its excessive activity can lead to AD and other skin diseases such as psoriasis and Netherton syndrome. This study proposes the use of *E. coli* SHuffle® T7 strain for the production of the LUP37C11 antibody in its scFv form. The SHuffle® T7 strain is recommended for producing recombinant proteins with disulfide bonds, as is the case with the target antibody. After production, the antibody will be purified, and its ability to recognize and inhibit hKLK7 will be assessed through ELISA assays and enzymatic kinetics.