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

Autor: Laureano AFS; Universidade Federal do ABC - Centro de Ciências Naturais e Humanas, São Bernardo do Campo, São Paulo, Brazil., Zani MB; Universidade Federal do ABC - Centro de Ciências Naturais e Humanas, São Bernardo do Campo, São Paulo, Brazil., Sant'Ana AM; Universidade Federal do ABC - Centro de Ciências Naturais e Humanas, São Bernardo do Campo, São Paulo, Brazil., Tognato RC; Universidade Federal do ABC - Centro de Ciências Naturais e Humanas, São Bernardo do Campo, São Paulo, Brazil., Lombello CB; Universidade Federal do ABC - Centro de Engenharia, Modelagem e Ciências Sociais aplicadas, São Bernardo do Campo, São Paulo, Brazil., do Nascimento MHM; Universidade Federal do ABC - Centro de Engenharia, Modelagem e Ciências Sociais aplicadas, São Bernardo do Campo, São Paulo, Brazil., Helmsing S; Technische Universität Braunschweig - Abteilung Biotechnologie, Institut für Biochemie, Biotechnologie und Bioinformatik, Braunschweig, Germany., Fühner V; Technische Universität Braunschweig - Abteilung Biotechnologie, Institut für Biochemie, Biotechnologie und Bioinformatik, Braunschweig, Germany., Hust M; Technische Universität Braunschweig - Abteilung Biotechnologie, Institut für Biochemie, Biotechnologie und Bioinformatik, Braunschweig, Germany., Puzer L; Universidade Federal do ABC - Centro de Ciências Naturais e Humanas, São Bernardo do Campo, São Paulo, Brazil. Electronic address: luciano.puzer@ufabc.edu.br.
Jazyk: angličtina
Zdroj: Bioorganic & medicinal chemistry letters [Bioorg Med Chem Lett] 2020 Dec 01; Vol. 30 (23), pp. 127626. Date of Electronic Publication: 2020 Oct 20.
DOI: 10.1016/j.bmcl.2020.127626
Abstrakt: 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 trypsin-like 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 naïve phage display library and characterized their inhibitory activities against KLK7. The scFv with the lowest IC 50 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 IC 50 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.
(Copyright © 2020 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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