Development of anti-matrix metalloproteinase-2 (MMP-2) nanobodies as potential therapeutic and diagnostic tools

Alessandro Marturano; Maarten L V Hendrickx; Emanuela Falcinelli; Manuela Sebastiano; Giuseppe Guglielmini; Gholamreza Hassanzadeh-Ghassabeh; Serge Muyldermans; Paul J Declerck; Paolo Gresele

doi:10.1016/j.nano.2019.102103

Development of anti-matrix metalloproteinase-2 (MMP-2) nanobodies as potential therapeutic and diagnostic tools

Nanomedicine. 2020 Feb:24:102103. doi: 10.1016/j.nano.2019.102103. Epub 2019 Oct 24.

Authors

Alessandro Marturano¹, Maarten L V Hendrickx², Emanuela Falcinelli¹, Manuela Sebastiano¹, Giuseppe Guglielmini¹, Gholamreza Hassanzadeh-Ghassabeh³, Serge Muyldermans⁴, Paul J Declerck², Paolo Gresele⁵

Affiliations

¹ Department of Medicine, Section of Internal and Cardiovascular Medicine, University of Perugia, Perugia, Italy.
² Laboratory for Therapeutic and Diagnostic Antibodies, KU, Leuven, Belgium.
³ Laboratory of Cellular and Molecular Immunology, Faculty of Sciences, Vrije Universiteit Brussel, Belgium; VIB-Nanobody Core, Brussels, Belgium.
⁴ Laboratory of Cellular and Molecular Immunology, Faculty of Sciences, Vrije Universiteit Brussel, Belgium.
⁵ Department of Medicine, Section of Internal and Cardiovascular Medicine, University of Perugia, Perugia, Italy. Electronic address: paolo.gresele@unipg.it.

PMID: 31669421
DOI: 10.1016/j.nano.2019.102103

Abstract

Matrix metalloproteinase-2 (MMP-2) is an endopeptidase involved in cardiovascular disease and cancer. To date, no highly selective MMP-2 inhibitors have been identified for potential use in humans. Aim of our work was to apply the nanobody technology to the generation of highly selective inhibitors of human MMP-2 and to assess their effects on platelet function and their applicability as conjugated nanobodies. We constructed a nanobody library after immunising an alpaca with human active MMP-2 and identified, after phage display and screening, one MMP-2 inhibitory nanobody (VHH-29), able to hinder the effects of MMP-2 on platelet activation, and one nanobody not inhibiting MMP-2 activity (VHH-136) which, chemically conjugated to a fluorescent probe, allowed the detection of human MMP-2 by flow-cytometry and immune-cytochemistry. In conclusion, we have generated and characterized two new nanotechnological molecular tools for human MMP-2 which represent promising agents for the study of MMP-2 in cardiovascular pathophysiology.

Keywords: Cancer; Nanobody; Platelet adhesion; Platelet priming; Thrombosis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Enzyme-Linked Immunosorbent Assay
Flow Cytometry*
HEK293 Cells
Humans
Matrix Metalloproteinase 2 / immunology*
Peptide Library*
Single-Domain Antibodies* / chemistry
Single-Domain Antibodies* / immunology

Substances

Peptide Library
Single-Domain Antibodies
MMP2 protein, human
Matrix Metalloproteinase 2