Journal: Nature Communications
Article Title: Supramolecular coiled-coil peptide platform for site-specific antibody drug conjugate engineering
doi: 10.1038/s41467-026-70094-y
Figure Lengend Snippet: A Stability of conventionally prepared AFCs via hinge cysteine conjugation, B via short 28-amino-acid-long coiled-coil peptides, C via long 35-amino-acid-long coiled-coil peptides, and D via long 35-amino-acid-long coiled-coil peptides with cysteine residue substitution in human plasma incubated at 37 °C. A – D Data are presented as mean ± s.d., n = 3 biological replicates. E Schematic illustration of ADC conjugation strategy and molecular structure of the linker-payload consisting of valine-citrulline (Val-Cit) cleavable portion and self-immolative p-aminobenzyl carbamate (PAB) moiety. F PK of unmodified trastuzumab (αHER2), unloaded ADC (αHER2-P3 5mer -cys), and fully assembled coiled-coil ADC (ADC) in female Balb/c. Blood was collected at the indicated time points, and the total amount of human IgG was quantified by indirect ELISA using a standard curve. Data are presented as mean ± s.d., n = 3 biological replicates for the trastuzumab group, n = 4 for ADC, and n = 2 for the unloaded ADC group. G Ex vivo organ biodistribution of AFC injected i.v. at a 10 mg/kg dose into SKOV-3 tumor-bearing BALB/c nude mice at different time points. Data are presented as mean ± s.d., n = 3 biological replicates. The total radiant efficiency was calculated using IVIS software, keeping the area of the ROI the same across all the samples. H Representative fluorescent picture of ex vivo organs post AFC injection. Color scale: min = 1.05 × 10 9 , max = 1.1 × 10 10 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$(\frac{p/\sec \frac{/{{cm}}^{2}}{{sr}}}{\mu W/{{cm}}^{2}})$$\end{document} ( p / sec / c m 2 s r μ W / c m 2 ) . I Orthotopic xenograft model of ErbB2/Her2-positive cancer study. Female NSG mice were engrafted with 2 million (2 M) SKOV-3 tumor cells by intraperitoneal injection (IP), were randomized on day 14, and treated with a single intravenous (IV) dose of PBS or trastuzumab antibody (10 mg/kg) or coiled-coil ADC (10 mg/kg) or high dose coiled-coil ADC (2xADC, 20 mg/kg) or disitamab vedotin (10 mg/kg), or trastuzumab vedotin (10 mg/kg), or disitamab antibody (10 mg/kg). All mice were euthanized at week 7 post-treatment, when PBS-treated mice exhibited severe hunching posture and signs of morbidity, to record the endpoint tumor volume and mass. J Average mouse body weight for selected treatment groups over the observation period. Data are presented as mean ± s.d., n = 5 biological replicates. K Exemplary images of harvested tumors at study termination. L Recorded tumor size, and M tumor mass. Data are presented as mean ± s.d., n = 5 biological replicates for Trastuzumab, Trastuzumab vedotin, ADC, 2xADC, and Disitamab treatment groups, n = 10 biological replicates for Disitamab vedotin, and n = 13 for PBS group. Statistical significance determined by the Mann–Whitney U test. Source data are provided as a Source Data file.
Article Snippet: Serial dilutions of native αHER2 (Human ErbB2/Her2 Antibody, Cat. # MAB9589-100, R&D Systems Inc) or antibodies fused to a receiving peptide (αHER2-P3) were prepared in blocking buffer (starting at 3 μg/mL).
Techniques: Conjugation Assay, Residue, Clinical Proteomics, Incubation, Indirect ELISA, Ex Vivo, Injection, Software, MANN-WHITNEY
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![A Stability of conventionally prepared AFCs via hinge cysteine conjugation, B via short 28-amino-acid-long coiled-coil peptides, C via long 35-amino-acid-long coiled-coil peptides, and D via long 35-amino-acid-long coiled-coil peptides with cysteine residue substitution in human plasma incubated at 37 °C. A – D Data are presented as mean ± s.d., n = 3 biological replicates. E Schematic illustration of ADC conjugation strategy and molecular structure of the linker-payload consisting of valine-citrulline (Val-Cit) cleavable portion and self-immolative p-aminobenzyl carbamate (PAB) moiety. F PK of unmodified trastuzumab (αHER2), unloaded ADC (αHER2-P3 5mer -cys), and fully assembled coiled-coil ADC (ADC) in female Balb/c. Blood was collected at the indicated time points, and the total amount of human IgG was quantified by indirect ELISA using a standard curve. Data are presented as mean ± s.d., n = 3 biological replicates for the trastuzumab group, n = 4 for ADC, and n = 2 for the unloaded ADC group. G Ex vivo organ biodistribution of AFC injected i.v. at a 10 mg/kg dose into SKOV-3 tumor-bearing BALB/c nude mice at different time points. Data are presented as mean ± s.d., n = 3 biological replicates. The total radiant efficiency was calculated using IVIS software, keeping the area of the ROI the same across all the samples. H Representative fluorescent picture of ex vivo organs post AFC injection. Color scale: min = 1.05 × 10 9 , max = 1.1 × 10 10 \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$(\frac{p/\sec \frac{/{{cm}}^{2}}{{sr}}}{\mu W/{{cm}}^{2}})$$\end{document} ( p / sec / c m 2 s r μ W / c m 2 ) . I Orthotopic xenograft model of ErbB2/Her2-positive cancer study. Female NSG mice were engrafted with 2 million (2 M) SKOV-3 tumor cells by intraperitoneal injection (IP), were randomized on day 14, and treated with a single intravenous (IV) dose of PBS or trastuzumab antibody (10 mg/kg) or coiled-coil ADC (10 mg/kg) or high dose coiled-coil ADC (2xADC, 20 mg/kg) or disitamab vedotin (10 mg/kg), or trastuzumab vedotin (10 mg/kg), or disitamab antibody (10 mg/kg). All mice were euthanized at week 7 post-treatment, when PBS-treated mice exhibited severe hunching posture and signs of morbidity, to record the endpoint tumor volume and mass. J Average mouse body weight for selected treatment groups over the observation period. Data are presented as mean ± s.d., n = 5 biological replicates. K Exemplary images of harvested tumors at study termination. L Recorded tumor size, and M tumor mass. Data are presented as mean ± s.d., n = 5 biological replicates for Trastuzumab, Trastuzumab vedotin, ADC, 2xADC, and Disitamab treatment groups, n = 10 biological replicates for Disitamab vedotin, and n = 13 for PBS group. Statistical significance determined by the Mann–Whitney U test. Source data are provided as a Source Data file.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_6297/pmc13096297/pmc13096297__41467_2026_70094_Fig4_HTML.jpg)