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Inhibition of cleavage of human complement component C5 and the R885H C5 variant by two distinct high affinity anti-C5 nanobodies
Open accessPreprint — published as 10.1016/j.jbc.2023.104956 · Paper page
Abstract
The human complement system plays a crucial role in immune defense. However, its erroneous activation contributes to many serious inflammatory diseases. Since most unwanted complement effector functions result from C5 cleavage, development of C5 inhibitors, such as clinically approved monoclonal antibody Eculizumab, are of great interest. In this study, we developed and characterized two anti-C5 nanobodies, UNbC5-1 and UNbC5-2. Using surface plasmon resonance (SPR), we determined a binding affinity of 120 pM for UNbC5-1 and 8 pM for UNbC5-2. Competition experiments determined that the two nanobodies recognize distinct epitopes on C5. Both nanobodies efficiently interfered with C5 cleavage in a human serum environment, as they prevented red blood cell lysis via membrane attack complexes (C5b-9) and the formation of chemoattractant C5a. The cryo-EM structure of UNbC5-1 and UNbC5-2 in complex with C5 revealed that the binding interfaces of UNbC5-1 and UNbC5-2 overlap with known complement inhibitors Eculizumab and RaCI3, respectively. UNbC5-1 binds to the MG7 domain of C5, facilitated by a hydrophobic core and polar interactions, and UNbC5-2 interacts with the C5d domain mostly by salt bridges and hydrogen bonds. Interestingly, UNbC5-1 potently binds and inhibits C5 R885H, a genetic variant of C5, that is not recognized by Eculizumab. Altogether, we identified and characterized two different, high affinity nanobodies against human C5. Both nanobodies could serve as diagnostic and/or research tools to detect C5 or inhibit C5 cleavage. Furthermore, the residues targeted by UNbC5-1 hold important information for therapeutic inhibition of different polymorphic variants of C5.
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Highly accurate protein structure prediction with AlphaFold
Jumper, J., Evans, R., Pritzel, A., Green, T., Figurnov, M., Ronneberger, O., Tunyasuvunakool, K., Bates, R., Žídek, A., Potapenko, A., et al. (2021) · Nature · doi:10.1038/s41586-021-03819-2 · Cited by 43,764
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Research works printed but not linked: 55
- 10.3389/fimmu.2019.00858 · DOI
- 2020, Role of complement 3 in renin generation during the differentiation of mesenchymal stem cells to smooth muscle cells, Am J Physiol Cell Physiol, 318, C981, 10.1152/ajpcell.00461.2019 · DOI
- 10.1016/j.smim.2019.101340 · DOI
- 10.3389/fimmu.2015.00001 · DOI
- 10.1124/pharmrev.120.000072 · DOI
- 10.3389/fimmu.2015.00001 · DOI
- 10.1084/jem.148.6.1722 · DOI
- 10.1073/pnas.0910565107 · DOI
- 2005, Complement Inhibitor of C5 Activation from the Soft Tick Ornithodoros moubata, The Journal of Immunology, 174, 2084, 10.4049/jimmunol.174.4.2084 · DOI
- 10.1073/pnas.1909973116 · DOI
- 10.1038/nsmb.3196 · DOI
- 10.1016/j.molimm.2018.06.008 · DOI
- 2018, Expanding horizons in complement drug discovery: challenges and emerging strategies, Semin Immunopathol, 40, 125, 10.1007/s00281-017-0655-8 · DOI
- 10.1038/nbt1344 · DOI · Paper page
- 10.1056/NEJMoa1311084 · DOI
- 2016, Structural Basis for Eculizumab-Mediated Inhibition of the Complement Terminal Pathway, The Journal of Immunology, 197, 337, 10.4049/jimmunol.1600280 · DOI
- 2016, Eculizumab epitope on complement C5: Progress towards a better understanding of the mechanism of action, Mol Immunol, 77, 126, 10.1016/j.molimm.2016.07.016 · DOI
- 2022, Multifaceted Activities of Seven Nanobodies against Complement C4b, The Journal of Immunology, 208, 2207, 10.4049/jimmunol.2100647 · DOI
- 10.1016/S0161-5890(02)00254-7 · DOI
- 10.1146/annurev.iy.04.040186.002443 · DOI
- 10.1038/nbt1344 · DOI · Paper page
- 2009, Eculizumab for paroxysmal nocturnal haemoglobinuria, The Lancet, 373, 759, 10.1016/S0140-6736(09)60001-5 · DOI
- 10.1038/ni.1625 · DOI
- 10.7554/eLife.62047 · DOI
- 1997, Structural definition of the C5a C terminus by two-dimensional nuclear magnetic resonance spectroscopy, Proteins: Structure, Function and Genetics, 28, 261, 10.1002/(SICI)1097-0134(199706)28:2<261::AID-PROT13>3.0.CO;2-G · DOI
- 10.3390/biom11020221 · DOI
- 10.1074/jbc.RA117.001179 · DOI
- 10.1038/ni.1756 · DOI
- 10.1038/emboj.2010.341 · DOI
- 10.3389/fimmu.2020.01022 · DOI
- 10.1074/jbc.RA119.012339 · DOI
- 2020, A Complement C3–Specific Nanobody for Modulation of the Alternative Cascade Identifies the C-Terminal Domain of C3b as Functional in C5 Convertase Activity, The Journal of Immunology, 205, 2287, 10.4049/jimmunol.2000752 · DOI
- 2020, An Ultrahigh-Affinity Complement C4b-Specific Nanobody Inhibits In Vivo Assembly of the Classical Pathway Proconvertase, The Journal of Immunology, 205, 1678, 10.4049/jimmunol.2000528 · DOI
- 2020, Recruitment of properdin by bi-specific nanobodies activates the alternative pathway of complement, Mol Immunol, 124, 200, 10.1016/j.molimm.2020.06.005 · DOI
- 10.3389/fimmu.2019.00858 · DOI
- 10.2967/jnumed.113.130617 · DOI
- 2017, Structural evaluation of a nanobody targeting complement receptor Vsig4 and its cross reactivity, Immunobiology, 222, 807, 10.1016/j.imbio.2016.11.008 · DOI
- 2018, Caplacizumab: First Global Approval, Drugs, 78, 1639, 10.1007/s40265-018-0989-0 · DOI
- 2021, Revealing the spatio-phenotypic patterning of cells in healthy and tumor tissues with mLSR-3D and STAPL-3D, Nat Biotechnol, 39, 1239, 10.1038/s41587-021-00926-3 · DOI
- 10.1073/pnas.1502609112 · DOI
- 10.1002/biot.201600357 · DOI
- 10.1016/S1567-5769(00)00039-4 · DOI
- 10.1111/imr.12465 · DOI
- 10.1111/cmi.12170 · DOI
- 10.1186/s12915-014-0111-3 · DOI
- 10.1371/journal.ppat.1009501 · DOI
- 10.1093/nar/19.15.4133 · DOI
- 2019, Vhh-photosensitizer conjugates for targeted photodynamic therapy of met-overexpressing tumor cells, Antibodies, 8, 1
- 10.1093/protein/gzr057 · DOI
- 10.1371/journal.pone.0179159 · DOI · Paper page
- 10.1002/PRO.3943 · DOI
- 10.1002/pro.3235 · DOI · Paper page
- 10.1107/S0907444904019158 · DOI · Paper page
- 10.1107/S2059798318006551 · DOI
- 2011, T-Coffee: A web server for the multiple sequence alignment of protein and RNA sequences using structural information and homology extension, Nucleic Acids Res, 39, 13, 10.1093/nar/gkr245 · DOI
Metadata
- Authors
- Struijf, E. M., De la O Becerra, K. I., Ruyken, M., van Oosterom, F., Siere, D. Y., Heesterbeek, D. A. C., Dolk, E., Heukers, R., Bardoel, B. W., Gros, P., & Rooijakkers, S. H. M.
- Affiliations
- Utrecht University (NL)
- Corresponding author
- Rooijakkers, Suzan H.M.
- ORCID
- Venue
- Journal of Biological Chemistry
- Publisher
- openRxiv
- Published
- 2023-02-22
- Volume
- 299
- Issue
- 8
- Pages
- 104956-104956
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC10374974
- OpenAlex
- W4381996895
- Cited by
- 15
- References
- 1
- Citations by year
- 2026: 6 · 2025: 7 · 2024: 2
- FWCI
- 1.64
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by-nc-nd
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by-nc-nd)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Utrecht University Repository (Utrecht University) (submittedVersion, other-oa)
- Funders
- European Commission (HORIZON2020)
- Consejo Nacional de Ciencia y Tecnología (604718, CVU 604718)
- Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWA.ID.17.036)
- Horizon 2020 Framework Programme
- European Research Council
- Horizon 2020 (101001937)
- H2020 European Research Council (787241)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Complement system in diseases
- Life Sciences › Immunology and Microbiology › Immunology › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Immunology and Microbiology › Immunology › Biomarkers in Disease Mechanisms
- Keywords
- complement component C5, nanobodies, C5a, membrane attack complex, eculizumab, epitope mapping, cryo-EM structure, complement system
- MeSH terms
- Antibodies, Monoclonal, Complement System Proteins, Complement Activation, Complement C5, Humans, Complement Membrane Attack Complex, Single-Domain Antibodies
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