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A selection and optimization strategy for single-domain antibodies targeting the PHF6 linear peptide within the tau intrinsically disordered protein

Mortelecque, Justine, Zejneli, Orgeta, Bégard, Séverine, Simões, Margarida C., ElHajjar, Lea, Nguyen, Marine, Cantrelle, François-Xavier, Hanoulle, Xavier, Rain, Jean-Christophe, Colin, Morvane, Gomes, Cláudio M., Buée, Luc, Landrieu, Isabelle, Danis, Clément, Dupré, Elian·2024·Journal of Biological Chemistry
DOI 10.1016/j.jbc.2024.107163PMC PMC11007443

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Figures

Figure 1 Selected mutations for VHH optimization.A, sequence of VHH Z70 and selected substitutions. CDR1, 2, and 3 sequences are represented in pink, blue, and red respectively. The three amino acids in bold italic are the most frequently substituted. When different substitutions exist that differ in frequency, the most frequent is in bold. B, overview of the mutants selected for further analysis. CDR, complementarity-defining region; VHH, variable heavy-chain of the heavy-chain–only antibody.
Figure 1 Selected mutations for VHH optimization.A, sequence of VHH Z70 and selected substitutions. CDR1, 2, and 3 sequences are represented in pink, blue, and red respectively. The three amino acids in bold italic are the most frequently substituted. When different substitutions exist that differ in frequency, the most frequent is in bold. B, overview of the mutants selected for further analysis. CDR, complementarity-defining region; VHH, variable heavy-chain of the heavy-chain–only antibody.CC BY · PubMed Central
Figure 2 Epitope verification of Z70 mutants.A, Western blot against tau using TAMRA-labeled VHHs. B, spectrum overlay of tau alone (in red), tau with Z70 (in blue, 1:1 M ratio), or tau with mutant 1 (in cyan, 1:1 M ratio). TAMRA, carboxytetramethylrhodamine; VHH, variable heavy-chain of the heavy-chain–only antibody.
Figure 2 Epitope verification of Z70 mutants.A, Western blot against tau using TAMRA-labeled VHHs. B, spectrum overlay of tau alone (in red), tau with Z70 (in blue, 1:1 M ratio), or tau with mutant 1 (in cyan, 1:1 M ratio). TAMRA, carboxytetramethylrhodamine; VHH, variable heavy-chain of the heavy-chain–only antibody.CC BY · PubMed Central
Figure 3 Epitope of Z70 and variant VHHs.A, schematic representation of the tau protein. N-terminal domain (1–163) is represented in red and may contain N1 and N2 inserted sequences depending on tau isoforms. The proline-rich domain (PRD) is subdivided in P1 and P2 regions. The microtubule binding domain consists of four partially repeated regions, R1 to R4 (in tau 4R). The R2 repeat is not present in tau 3R. C-ter is for the C-terminal domain. B, normalized intensity ratios I/I0 of corresponding resonances in the two-dimensional spectra of tau with equimolar quantity of Z70, mutant 1, 3, or 20 (I) or free in solution (I0) for residues along the tau sequence.
Figure 3 Epitope of Z70 and variant VHHs.A, schematic representation of the tau protein. N-terminal domain (1–163) is represented in red and may contain N1 and N2 inserted sequences depending on tau isoforms. The proline-rich domain (PRD) is subdivided in P1 and P2 regions. The microtubule binding domain consists of four partially repeated regions, R1 to R4 (in tau 4R). The R2 repeat is not present in tau 3R. C-ter is for the C-terminal domain. B, normalized intensity ratios I/I0 of corresponding resonances in the two-dimensional spectra of tau with equimolar quantity of Z70, mutant 1, 3, or 20 (I) or free in solution (I0) for residues along the tau sequence.CC BY · PubMed Central
Figure 4 Comparison of Z70, mutant 1, 3, and 20 structures. Superposition of Z70 (in green, PDB 7qcq), mutant 1 (in gold, PDB 8opi), mutant 3 (in pale green, PDB 8pii), and mutant 20 (in orange, PDB 8op0). Their CDR1 are in pink, CDR2 in blue, and CDR3 in purple. Tau PHF6 peptide is colored cyan. CDR, complementarity-defining region; PDB, Protein Data Bank.
Figure 4 Comparison of Z70, mutant 1, 3, and 20 structures. Superposition of Z70 (in green, PDB 7qcq), mutant 1 (in gold, PDB 8opi), mutant 3 (in pale green, PDB 8pii), and mutant 20 (in orange, PDB 8op0). Their CDR1 are in pink, CDR2 in blue, and CDR3 in purple. Tau PHF6 peptide is colored cyan. CDR, complementarity-defining region; PDB, Protein Data Bank.CC BY · PubMed Central
Figure 5 Comparison of Z70, mutant 1, 3, and 20 electron density maps. Z70, mutants 1, 3, and 20 are represented in green, gold, pale green and orange, respectively. Tau PHF6 peptide is colored cyan. A, environment of the peptide in all three structures remains unchanged. B, electron density (2fo-fc map contoured at 1 σ), shown as mesh representation, corresponding to the region surrounding residues 115 to 117 (left) and 46 to 48 (right) in VHH Z70 (top), mutant 1 (middle) and mutant 20 (bottom), respectively, showing the appearance of density for residue 115 from Z70 to mutant 20. A salt bridge is shown between K47 and E115 of mutant 20 in red dashes. VHH, variable heavy-chain of the heavy-chain–only antibody.
Figure 5 Comparison of Z70, mutant 1, 3, and 20 electron density maps. Z70, mutants 1, 3, and 20 are represented in green, gold, pale green and orange, respectively. Tau PHF6 peptide is colored cyan. A, environment of the peptide in all three structures remains unchanged. B, electron density (2fo-fc map contoured at 1 σ), shown as mesh representation, corresponding to the region surrounding residues 115 to 117 (left) and 46 to 48 (right) in VHH Z70 (top), mutant 1 (middle) and mutant 20 (bottom), respectively, showing the appearance of density for residue 115 from Z70 to mutant 20. A salt bridge is shown between K47 and E115 of mutant 20 in red dashes. VHH, variable heavy-chain of the heavy-chain–only antibody.CC BY · PubMed Central
Figure 6 Conformational stabilization of mutant 20.A, cartoon representation of mutant 20 showing the salt bridge between residues K47 and E115 as red dashes. The framework region is colored orange, CDR 1 is represented in pink, CDR2 in blue, and CDR3 in purple, while the PHF6 peptide is colored cyan. B, superimposition of 1H 15N HSQC spectra of 15N labeled mutant 20 in blue and VHH Z70 in red showing the detection of additional resonances in the spectrum of mutant 20 indicating a change in the conformational dynamics that could be favored by the K47-E115 salt bridge. CDR, complementarity-defining region; HSQC, heteronuclear single quantum coherence; VHH, variable heavy-chain of the heavy-chain–only antibody.
Figure 6 Conformational stabilization of mutant 20.A, cartoon representation of mutant 20 showing the salt bridge between residues K47 and E115 as red dashes. The framework region is colored orange, CDR 1 is represented in pink, CDR2 in blue, and CDR3 in purple, while the PHF6 peptide is colored cyan. B, superimposition of 1H 15N HSQC spectra of 15N labeled mutant 20 in blue and VHH Z70 in red showing the detection of additional resonances in the spectrum of mutant 20 indicating a change in the conformational dynamics that could be favored by the K47-E115 salt bridge. CDR, complementarity-defining region; HSQC, heteronuclear single quantum coherence; VHH, variable heavy-chain of the heavy-chain–only antibody.CC BY · PubMed Central
Figure 7 Thermal stabilities of the different VHHsin NMR buffer. VHH, variable heavy-chain of the heavy-chain–only antibody.
Figure 7 Thermal stabilities of the different VHHsin NMR buffer. VHH, variable heavy-chain of the heavy-chain–only antibody.CC BY · PubMed Central
Figure 8 In-cell stabilities of the VHHs.A, percentage of mCherry-positive HEK293 cells showing aggregates (puncta) of VHH-mCherry 48 h after transfection. B, Pearson plot of the percentage of cells with aggregates in the self-aggregation assay versus melting temperatures in NMR buffer. VHH, variable heavy-chain of the heavy-chain–only antibody.
Figure 8 In-cell stabilities of the VHHs.A, percentage of mCherry-positive HEK293 cells showing aggregates (puncta) of VHH-mCherry 48 h after transfection. B, Pearson plot of the percentage of cells with aggregates in the self-aggregation assay versus melting temperatures in NMR buffer. VHH, variable heavy-chain of the heavy-chain–only antibody.CC BY · PubMed Central
Figure 9 In vitro inhibition of tau aggregation by the VHHs.A, inhibition of tau-K18 aggregation by Z70 at VHH:Tau-K18 M ratios between 0.1 and 0.4 (n = 4) B, inhibition efficiency of different Z70-derived VHHs over tau aggregation tested at a molar ratio of 0.5:1 VHH:tau. Positive and negative controls correspond to tau in the presence of heparin (tau +) and the absence of heparin (tau −), respectively. The other conditions correspond to tau in the presence of heparin and the different VHHs at a VHH:tau ratio of 0.5:1. The percentage of aggregation corresponds to the fluorescence intensity ratio between tau in the presence of a VHH and tau alone (tau +) during the aggregation process. Data are represented as mean ± SEM, the mean value is written above the bars (n = 3). VHH, variable heavy-chain of the heavy-chain–only antibody.
Figure 9 In vitro inhibition of tau aggregation by the VHHs.A, inhibition of tau-K18 aggregation by Z70 at VHH:Tau-K18 M ratios between 0.1 and 0.4 (n = 4) B, inhibition efficiency of different Z70-derived VHHs over tau aggregation tested at a molar ratio of 0.5:1 VHH:tau. Positive and negative controls correspond to tau in the presence of heparin (tau +) and the absence of heparin (tau −), respectively. The other conditions correspond to tau in the presence of heparin and the different VHHs at a VHH:tau ratio of 0.5:1. The percentage of aggregation corresponds to the fluorescence intensity ratio between tau in the presence of a VHH and tau alone (tau +) during the aggregation process. Data are represented as mean ± SEM, the mean value is written above the bars (n = 3). VHH, variable heavy-chain of the heavy-chain–only antibody.CC BY · PubMed Central
Figure 10 In cellulo inhibition of tau seeding by the VHHs. The percentage of FRET-positive cells, corresponding to cells with aggregated tau, is given for the mCherry-gated population, corresponding to transfected cells. Values are normalized relative to the experiment with F8-2 VHH-mCherry. Data are represented as mean ± SEM, the mean value is given above the bars and individual points are plotted. Statistical comparison between each VHH and F8-2 was carried out using post hoc Dunn’s test after Kruskall–Wallis test with p = 2.9∗10-5, ∗∗∗ is p < 0.001, ∗∗ p < 0.01, and – is not significant. VHH, variable heavy-chain of the heavy-chain–only antibody.
Figure 10 In cellulo inhibition of tau seeding by the VHHs. The percentage of FRET-positive cells, corresponding to cells with aggregated tau, is given for the mCherry-gated population, corresponding to transfected cells. Values are normalized relative to the experiment with F8-2 VHH-mCherry. Data are represented as mean ± SEM, the mean value is given above the bars and individual points are plotted. Statistical comparison between each VHH and F8-2 was carried out using post hoc Dunn’s test after Kruskall–Wallis test with p = 2.9∗10-5, ∗∗∗ is p < 0.001, ∗∗ p < 0.01, and – is not significant. VHH, variable heavy-chain of the heavy-chain–only antibody.CC BY · PubMed Central
Figure 11 Pearson correlation plots.A, mean values of percentage of tau seeding in HEK293 tau repeat domain (RD) P301S FRET Biosensor seeding reporter cells (cellular seeding assay) versus mean values of in vitro aggregation assay with a ratio of 0.5 VHH for 1 tau. Dashed line is omitting the outlier mutant 9. B, mean values of percentage of tau seeding in HEK293 tau repeat domain (RD) P301S FRET biosensor seeding reporter cells (cellular seeding assay) versus percentage of cells showing punctate aggregates in HEK293 cells. VHH, variable heavy-chain of the heavy-chain–only antibody.
Figure 11 Pearson correlation plots.A, mean values of percentage of tau seeding in HEK293 tau repeat domain (RD) P301S FRET Biosensor seeding reporter cells (cellular seeding assay) versus mean values of in vitro aggregation assay with a ratio of 0.5 VHH for 1 tau. Dashed line is omitting the outlier mutant 9. B, mean values of percentage of tau seeding in HEK293 tau repeat domain (RD) P301S FRET biosensor seeding reporter cells (cellular seeding assay) versus percentage of cells showing punctate aggregates in HEK293 cells. VHH, variable heavy-chain of the heavy-chain–only antibody.CC BY · PubMed Central

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Research works printed but not linked: 71

  1. Hamers-Casterman, 1993, Naturally occurring antibodies devoid of light chains, Nature, 363, 446, 10.1038/363446a0 · DOI · Paper page
  2. Jovčevska, 2020, The therapeutic potential of nanobodies, BioDrugs, 34, 11, 10.1007/s40259-019-00392-z · DOI · Paper page
  3. Pleiner, 2015, Nanobodies: site-specific labeling for super-resolution imaging, rapid epitope-mapping and native protein complex isolation, eLife, 4, 10.7554/eLife.11349 · DOI
  4. Barakat, 2022, Nanobodies as molecular imaging probes, Free Radic. Biol. Med., 182, 260, 10.1016/j.freeradbiomed.2022.02.031 · DOI
  5. Jiang, 2023, Single-domain antibody–based noninvasive in vivo imaging of α-synuclein or tau pathology, Sci. Adv., 9, 10.1126/sciadv.adf3775 · DOI
  6. Peyvandi, 2016, Caplacizumab for acquired thrombotic thrombocytopenic purpura, N. Engl. J. Med., 374, 511, 10.1056/NEJMoa1505533 · DOI
  7. Takeuchi, 2022, Phase II/III results of a trial of anti–tumor necrosis factor multivalent NANOBODY compound ozoralizumab in patients with rheumatoid arthritis, Arthritis Rheumatol., 74, 1776, 10.1002/art.42273 · DOI
  8. Ishiwatari-Ogata, 2022, Ozoralizumab, a humanized anti-TNFα NANOBODY® compound, exhibits efficacy not only at the onset of arthritis in a human TNF transgenic mouse but also during secondary failure of administration of an anti-TNFα IgG, Front. Immunol., 13, 10.3389/fimmu.2022.853008 · DOI
  9. Messer, 2020, Optimizing intracellular antibodies (intrabodies/nanobodies) to treat neurodegenerative disorders, Neurobiol. Dis., 134, 10.1016/j.nbd.2019.104619 · DOI
  10. Zheng, 2022, Applications of nanobodies in brain diseases, Front. Immunol., 13, 10.3389/fimmu.2022.978513 · DOI
  11. Tsitokana, 2023, Targeting the brain with single-domain antibodies: greater potential than stated so far?, Int. J. Mol. Sci., 24, 2632, 10.3390/ijms24032632 · DOI
  12. Frenken, 2000, Isolation of antigen specific Llama VHH antibody fragments and their high level secretion by Saccharomyces cerevisiae, J. Biotechnol., 78, 11, 10.1016/S0168-1656(99)00228-X · DOI
  13. Moutel, 2016, NaLi-H1: a universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies, Elife, 5, 10.7554/eLife.16228 · DOI
  14. Desmyter, 2015, Camelid nanobodies: killing two birds with one stone, Curr. Opin. Struct. Biol., 32, 1, 10.1016/j.sbi.2015.01.001 · DOI
  15. Muyldermans, 2021, Applications of nanobodies, Annu. Rev. Anim. Biosci., 9, 401, 10.1146/annurev-animal-021419-083831 · DOI
  16. Danis, 2022, Inhibition of Tau seeding by targeting Tau nucleation core within neurons with a single domain antibody fragment, Mol. Ther., 30, 1484, 10.1016/j.ymthe.2022.01.009 · DOI · Paper page
  17. Guilliams, 2013, Nanobodies raised against monomeric α-synuclein distinguish between fibrils at different maturation stages, J. Mol. Biol., 425, 2397, 10.1016/j.jmb.2013.01.040 · DOI
  18. Lin, 2021, Computational maturation of a single-domain antibody against Aβ42 aggregation, Chem. Sci., 12, 13940, 10.1039/D1SC03898B · DOI
  19. Abskharon, 2014, Probing the N-terminal β-sheet conversion in the crystal structure of the human prion protein bound to a nanobody, J. Am. Chem. Soc., 136, 937, 10.1021/ja407527p · DOI
  20. Dupré, 2019, Single domain antibody fragments as new tools for the detection of neuronal tau protein in cells and in mice studies, ACS Chem. Neurosci., 10, 3997, 10.1021/acschemneuro.9b00217 · DOI
  21. Chan, 2008, Engineering a camelid antibody fragment that binds to the active site of human lysozyme and inhibits its conversion into amyloid fibrils, Biochemistry, 47, 11041, 10.1021/bi8005797 · DOI
  22. Abskharon, 2023, Structure-based design of nanobodies that inhibit seeding of Alzheimer’s patient–extracted tau fibrils, Proc. Natl. Acad. Sci., 120, 10.1073/pnas.2300258120 · DOI
  23. Fichou, 2019, The elusive tau molecular structures: can we translate the recent breakthroughs into new targets for intervention?, Acta Neuropathol. Commun., 7, 31, 10.1186/s40478-019-0682-x · DOI
  24. Fitzpatrick, 2017, Cryo-EM structures of tau filaments from Alzheimer’s disease, Nature, 547, 185, 10.1038/nature23002 · DOI · Paper page
  25. Scheres, 2023, Molecular pathology of neurodegenerative diseases by cryo-EM of amyloids, Nature, 621, 701, 10.1038/s41586-023-06437-2 · DOI
  26. Mudher, 2017, What is the evidence that tau pathology spreads through prion-like propagation?, Acta Neuropathol. Commun., 5, 99, 10.1186/s40478-017-0488-7 · DOI · Paper page
  27. Goedert, 2020, Chapter Ten - tau proteinopathies and the prion concept, Prog. Mol. Biol. Transl. Sci., 175, 239, 10.1016/bs.pmbts.2020.08.003 · DOI · Paper page
  28. Goedert, 2010, The propagation of prion-like protein inclusions in neurodegenerative diseases, Trends Neurosci., 33, 317, 10.1016/j.tins.2010.04.003 · DOI
  29. Franzmeier, 2020, Functional brain architecture is associated with the rate of tau accumulation in Alzheimer’s disease, Nat. Commun., 11, 347, 10.1038/s41467-019-14159-1 · DOI
  30. Franzmeier, 2022, Tau deposition patterns are associated with functional connectivity in primary tauopathies, Nat. Commun., 13, 1362, 10.1038/s41467-022-28896-3 · DOI
  31. Liu, 2018, Expression of single-domain antibody in different systems, Appl. Microbiol. Biotechnol., 102, 539, 10.1007/s00253-017-8644-3 · DOI
  32. Chao, 2022, Highly expressed soluble recombinant anti-GFP VHHs in Escherichia coli via optimized signal peptides, strains, and inducers, Front. Mol. Biosci., 9, 10.3389/fmolb.2022.848829 · DOI
  33. Sulea, 2022, Structure-based dual affinity optimization of a SARS-CoV-1/2 cross-reactive single-domain antibody, PLoS One, 17, 10.1371/journal.pone.0266250 · DOI
  34. Goldman, 2017, Enhancing stability of camelid and shark single domain antibodies: an overview, Front. Immunol., 8, 865, 10.3389/fimmu.2017.00865 · DOI
  35. Dingus, 2022, A general approach for stabilizing nanobodies for intracellular expression, Elife, 11, 10.7554/eLife.68253 · DOI
  36. Wagner, 2021, Nanobodies – little helpers unravelling intracellular signaling, Free Radic. Biol. Med., 176, 46, 10.1016/j.freeradbiomed.2021.09.005 · DOI
  37. Kvam, 2010, Physico-chemical determinants of soluble intrabody expression in mammalian cell cytoplasm, Protein Eng. Des. Sel., 23, 489, 10.1093/protein/gzq022 · DOI
  38. Fisher, 2009, Efficient isolation of soluble intracellular single-chain antibodies using the twin-arginine translocation machinery, J. Mol. Biol., 385, 299, 10.1016/j.jmb.2008.10.051 · DOI
  39. da Silva, 2004, Camelized rabbit-derived VH single-domain intrabodies against vif strongly neutralize HIV-1 infectivity, J. Mol. Biol., 340, 525, 10.1016/j.jmb.2004.04.062 · DOI
  40. Martineau, 1998, Expression of an antibody fragment at high levels in the bacterial cytoplasm, J. Mol. Biol., 280, 117, 10.1006/jmbi.1998.1840 · DOI
  41. Strube, 2004, Enhanced intracellular stability of sFv–Fc fusion intrabodies, Methods, 34, 179, 10.1016/j.ymeth.2004.04.003 · DOI
  42. Shaki-Loewenstein, 2005, A universal strategy for stable intracellular antibodies, J. Immunological Methods, 303, 19, 10.1016/j.jim.2005.05.004 · DOI
  43. Danis, 2019, Nuclear magnetic resonance spectroscopy insights into tau structure in solution: impact of post-translational modifications, Adv. Exp. Med. Biol., 1184, 35, 10.1007/978-981-32-9358-8_3 · DOI
  44. Rennella, 2023, Exploiting conformational dynamics to modulate the function of designed proteins, Proc. Natl. Acad. Sci. U. S. A., 120, 10.1073/pnas.2303149120 · DOI
  45. Alderson, 2021, NMR spectroscopy captures the essential role of dynamics in regulating biomolecular function, Cell, 184, 577, 10.1016/j.cell.2020.12.034 · DOI
  46. Shammas, 2015, A mechanistic model of tau amyloid aggregation based on direct observation of oligomers, Nat. Commun., 6, 10.1038/ncomms8025 · DOI
  47. von Bergen, 2000, Assembly of tau protein into Alzheimer paired helical filaments depends on a local sequence motif ((306)VQIVYK(311)) forming beta structure, Proc. Natl. Acad. Sci. U. S. A., 97, 5129, 10.1073/pnas.97.10.5129 · DOI · Paper page
  48. Lövestam, 2023, Disease-specific tau filaments assemble via polymorphic intermediates, Nature, 625, 119, 10.1038/s41586-023-06788-w · DOI
  49. Holmes, 2014, Proteopathic tau seeding predicts tauopathy in vivo, Proc. Natl. Acad. Sci. U. S. A., 111, E4376, 10.1073/pnas.1411649111 · DOI · Paper page
  50. Luginbühl, 2006, Directed evolution of an anti-prion protein scFv fragment to an affinity of 1 pM and its structural interpretation, J. Mol. Biol., 363, 75, 10.1016/j.jmb.2006.07.027 · DOI
  51. Van Roy, 2015, The preclinical pharmacology of the high affinity anti-IL-6R Nanobody® ALX-0061 supports its clinical development in rheumatoid arthritis, Arthritis Res. Ther., 17, 135, 10.1186/s13075-015-0651-0 · DOI
  52. Braun, 2016, Peptides in headlock – a novel high-affinity and versatile peptide-binding nanobody for proteomics and microscopy, Sci. Rep., 6, 10.1038/srep19211 · DOI
  53. Sheinerman, 2002, On the role of electrostatic interactions in the design of protein-protein interfaces, J. Mol. Biol., 318, 161, 10.1016/S0022-2836(02)00030-X · DOI
  54. Selzer, 2000, Rational design of faster associating and tighter binding protein complexes, Nat. Struct. Biol., 7, 537, 10.1038/76744 · DOI
  55. Govaert, 2012, Dual beneficial effect of interloop disulfide bond for single domain antibody fragments, J. Biol. Chem., 287, 1970, 10.1074/jbc.M111.242818 · DOI
  56. Kunz, 2017, Exploiting sequence and stability information for directing nanobody stability engineering, Biochim. Biophys. Acta Gen. Subj., 1861, 2196, 10.1016/j.bbagen.2017.06.014 · DOI
  57. Cadwell, 1992, Randomization of genes by PCR mutagenesis, PCR Methods Appl., 2, 28, 10.1101/gr.2.1.28 · DOI
  58. Vojtek, 1995, Ras-Raf interaction: two-hybrid analysis, Meth. Enzymol., 255, 331, 10.1016/S0076-6879(95)55036-4 · DOI
  59. Fromont-Racine, 1997, Toward a functional analysis of the yeast genome through exhaustive two-hybrid screens, Nat. Genet., 16, 277, 10.1038/ng0797-277 · DOI
  60. Danis, 2016, Nuclear magnetic resonance spectroscopy for the identification of multiple phosphorylations of intrinsically disordered proteins, J. Vis. Exp., 118
  61. Moreira, 2021, Dynamic interactions and Ca2+-binding modulate the holdase-type chaperone activity of S100B preventing tau aggregation and seeding, Nat. Commun., 12, 6292, 10.1038/s41467-021-26584-2 · DOI · Paper page
  62. Lee, 2021, POKY: a software suite for multidimensional NMR and 3D structure calculation of biomolecules, Bioinformatics, 37, 3041, 10.1093/bioinformatics/btab180 · DOI
  63. Hunter, 2007, Matplotlib: a 2D graphics environment, Comput. Sci. Eng., 9, 90, 10.1109/MCSE.2007.55 · DOI · Paper page
  64. Karlsson, 2006, Analyzing a kinetic titration series using affinity biosensors, Anal. Biochem., 349, 136, 10.1016/j.ab.2005.09.034 · DOI · Paper page
  65. Vagin, 2010, Molecular replacement with MOLREP, Acta Crystallogr. D Biol. Crystallogr., 66, 22, 10.1107/S0907444909042589 · DOI
  66. Murshudov, 2011, REFMAC5 for the refinement of macromolecular crystal structures, Acta Crystallogr. D Biol. Crystallogr., 67, 355, 10.1107/S0907444911001314 · DOI
  67. Emsley, 2010, Features and development of coot, Acta Crystallogr. D Biol. Crystallogr., 66, 486, 10.1107/S0907444910007493 · DOI · Paper page
  68. Waskom, 2021, seaborn: statistical data visualization, J. Open Source Softw., 6, 3021, 10.21105/joss.03021 · DOI · Paper page
  69. Banning, 2010, A flow cytometry-based FRET assay to identify and analyse protein-protein interactions in living cells, PLoS One, 5, 10.1371/journal.pone.0009344 · DOI
  70. White, 2021, FlowKit: a Python toolkit for integrated manual and automated cytometry analysis workflows, Front. Immunol., 12, 10.3389/fimmu.2021.768541 · DOI
  71. Virtanen, 2020, SciPy 1.0: fundamental algorithms for scientific computing in Python, Nat. Methods, 17, 261, 10.1038/s41592-019-0686-2 · DOI · Paper page

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Metadata

Authors
Mortelecque, J., Zejneli, O., Bégard, S., Simões, M. C., ElHajjar, L., Nguyen, M., Cantrelle, F.-X., Hanoulle, X., Rain, J.-C., Colin, M., Gomes, C. M., Buée, L., Landrieu, I., Danis, C., & Dupré, E.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2024-04
Volume
300
Issue
4
Pages
107163
ISSN
0021-9258
DOI
10.1016/j.jbc.2024.107163
PMC
PMC11007443
Cited by
12
References
1
Language
en
Licence
Funders
  • Fundação para a Ciência e a Tecnologia (GA 101079147, UIDP/MULTI/04046/2020, UIDB/04046/2020, 2022.11023)
  • Institute Pasteur De Lille
  • French National Research Agency (ANR-18-CE44-0016, ANR-22-CE92-0061, ANR-11-LABX-01)
  • Campus France
  • Labex
  • Centre National de la Recherche Scientifique
  • European Commission

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CoChat. (2026). Citation check: A selection and optimization strategy for single-domain antibodies targeting the PHF6 linear peptide within the tau intrinsically disordered protein. https://app.cochat.ai/papers/10.1016/j.jbc.2024.107163
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Mortelecque, J., Zejneli, O., Bégard, S., Simões, M. C., ElHajjar, L., Nguyen, M., Cantrelle, F.-X., Hanoulle, X., Rain, J.-C., Colin, M., Gomes, C. M., Buée, L., Landrieu, I., Danis, C., & Dupré, E. (2024). A selection and optimization strategy for single-domain antibodies targeting the PHF6 linear peptide within the tau intrinsically disordered protein. Journal of Biological Chemistry. https://doi.org/10.1016/j.jbc.2024.107163

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Bibliographic data: Semantic Scholar, OpenAlex, Crossref.

Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).

Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.