A potent complement factor C3-specific nanobody inhibiting multiple functions in the alternative pathway of human and murine complement

The Journal of Biological Chemistry
Rasmus K JensenGregers R Andersen

Abstract

The complement system is a complex, carefully regulated proteolytic cascade for which suppression of aberrant activation is of increasing clinical relevance, and inhibition of the complement alternative pathway is a subject of intense research. Here, we describe the nanobody hC3Nb1 that binds to multiple functional states of C3 with subnanomolar affinity. The nanobody causes a complete shutdown of alternative pathway activity in human and murine serum when present in concentrations comparable with that of C3, and hC3Nb1 is shown to prevent proconvertase assembly, as well as binding of the C3 substrate to C3 convertases. Our crystal structure of the C3b-hC3Nb1 complex and functional experiments demonstrate that proconvertase formation is blocked by steric hindrance between the nanobody and an Asn-linked glycan on complement factor B. In addition, hC3Nb1 is shown to prevent factor H binding to C3b, rationalizing its inhibition of factor I activity. Our results identify hC3Nb1 as a versatile, inexpensive, and powerful inhibitor of the alternative pathway in both human and murine in vitro model systems of complement activation.

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Citations

May 8, 2020·The Journal of Biological Chemistry·Henrik PedersenGregers R Andersen
Sep 12, 2019·Frontiers in Immunology·Dennis V PedersenGregers R Andersen
Jun 26, 2020·Immunology·Jens Magnus Bernth JensenSteffen Thiel
Aug 10, 2020·The Journal of Immunology : Official Journal of the American Association of Immunologists·Alessandra ZarantonelloGregers R Andersen
Sep 18, 2020·The Journal of Immunology : Official Journal of the American Association of Immunologists·Henrik PedersenGregers R Andersen
Dec 20, 2020·Immunology·Jens Magnus Bernth JensenSteffen Thiel
Jan 23, 2021·ELife·Dennis V PedersenGregers R Andersen
Feb 2, 2021·Science Advances·Rasmus PihlSteffen Thiel
Nov 19, 2020·Medical Microbiology and Immunology·Mads LausenSvend Birkelund
Jun 13, 2021·The Journal of Immunology : Official Journal of the American Association of Immunologists·Rasmus K JensenGregers R Andersen
Jun 20, 2020·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Xuedong HeCalvin Yu-Chian Chen

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