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Expression and inhibitory characterization of scFv antibody fragments against kallikrein 7

Grant number: 19/11045-1
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): November 01, 2019
Effective date (End): July 31, 2021
Field of knowledge:Biological Sciences - Biochemistry - Molecular Biology
Principal Investigator:Luciano Puzer
Grantee:Rafael Cerioni Tognato
Host Institution: Centro de Ciências Naturais e Humanas (CCNH). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil

Abstract

Protease inhibitors are molecules capable of directly or indirectly affecting the active site of proteolytic enzymes, limiting or eliminating their ability to catalyze the cleavage of peptide bonds. These molecules can still act selectively or specifically, endogenously controlling the most diverse physiological processes. In addition, protease inhibitors also represent a considerable share of the drug market and are used in therapeutic procedures where proteases play some relevant role in pathological development. In recent years our research group has been active in the development of inhibitors for proteases, specifically against human tissue kallikreins (KLK), which comprise a family of 15 serine proteases (KLK1-KLK15) distributed in the most varied tissues of the human body , and which present several physiological and pathological functions. Recombinant antibodies of the sc-Fv-Fc type against (KLK7) were previously selected, expressed in HEK 293 human lineage and characterized as to their ability to inhibit the action of protease. The best antibodies were still used in formulations for treatment of skin diseases using a hydrogel drug delivery system. Thus, in this project, we propose the expression and inhibitory characterization of scFv-like antibody fragments. Our hypothesis is that these molecules will also have the ability to inhibit KLK7, but with a much lower production cost, since they are expressed in bacteria. In addition, these fragments, being smaller, can be more easily encapsulated in the hydrogels for drug delivery.

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