Generation of recombinant antibodies against human tissue kallikrein 7 to treat skin diseases

Autor(es)

Laureano, Ana Flavia S. ^[1] ; Zani, Marcelo B. ^[1] ; Sant'Ana, Aquiles M. ^[1] ; Tognato, Rafael C. ^[1] ; Lombello, Christiane B. ^[2] ; do Nascimento, Monica Helena M. ^[2] ; Helmsing, Saskia ^[3] ; Fuehner, Viola ^[3] ; Hust, Michael ^[3] ; Puzer, Luciano ^[1]

Afiliação do(s) autor(es)

^[1] Univ Fed ABC, Ctr Ciencias Nat & Humanas, Sao Bernardo Do Campo, SP - Brazil

^[2] Univ Fed ABC, Ctr Engn Modelagem & Ciencias Sociais Aplicadas, Sao Bernardo Do Campo, SP - Brazil

^[3] Tech Univ Carolo Wilhelmina Braunschweig, Inst Biochem Biotechnol & Bioinformat, Abt Biotechnol, Braunschweig - Germany

• Tipo de documento: Artigo Científico
• Fonte: Bioorganic & Medicinal Chemistry Letters; v. 30, n. 23 DEC 1 2020.
• Citações Web of Science: 0

Resumo

Human tissue kallikreins (KLKs) constitute a family of 15 serine proteases that are distributed in various tissues and implicated in several pathological disorders. KLK7 is an unusual serine protease that presents both trypsinlike and chymotrypsin-like specificity and appears to be upregulated in pathologies that are related to skin desquamation processes, such as atopic dermatitis, psoriasis and Netherton syndrome. In recent years, various groups have worked to develop specific inhibitors for this enzyme, as KLK7 represents a potential target for new therapeutic procedures for diseases related to skin desquamation processes. In this work, we selected nine different single-chain variable fragment antibodies (scFv) from a human naive phage display library and characterized their inhibitory activities against KLK7. The scFv with the lowest IC50 against KLK7 was affinity maturated, which resulted in the generation of four new scFv-specific antibodies for the target protease. These new antibodies were expressed in the scFv-Fc format in HEK293-6E cells, and the characterization of their inhibitory activities against KLK7 showed that three of them presented IC50 values lower than that of the original antibody. The cytotoxicity analysis of these recombinant antibodies demonstrated that they can be safely used in a cellular model. In conclusion, our research showed that in our case, a phage-display methodology in combination with enzymology assays can be a very suitable tool for the development of inhibitors for KLKs, suggesting a new strategy to identify therapeutic protease inhibitors for diseases related to uncontrolled kallikrein activity. (AU)

• Processo FAPESP: 16/21441-3 - Desenvolvimento de Inibidores para as Calicreínas Tecidas Humanas
• Beneficiário: Luciano Puzer
• Modalidade de apoio: Auxílio à Pesquisa - Regular