Journal: Scientific reports

Article Title: GC1126A, a novel ADAMTS13 mutein, evades autoantibodies in immune-mediated thrombotic thrombocytopenic purpura.

doi: 10.1038/s41598-024-80674-x

Figure Lengend Snippet: Fig. 1. The process and outcomes of candidate screening and the pharmacokinetic profile of candidates. (a) The diagram shows the binding domains of anti-ADAMTS13 neutralizing antibodies (Nabs). (b) The strategy and process of candidate screening. (c) The specific activity of selected candidates in the media following transient expression (n = 1 or 2). (d) The relative residual activity of selected candidates in the presence of 9 Nabs (n = 1 or 2). (e) The plasma concentration profiles after the intravenous administration of the selected candidates or controls (MDTCS or MDTCS-Fc) at the dose level of 160 IU/kg. Each point represents the mean and standard error of the mean (n = 4 mice per time point).

Article Snippet: The recombinant human ADAMTS13 (full-length) protein (rhADAMTS13 FL; R&D Systems, Minnesota, USA) served as the antigen.

Techniques: Binding Assay, Activity Assay, Expressing, Clinical Proteomics, Concentration Assay

Journal: Scientific reports

Article Title: GC1126A, a novel ADAMTS13 mutein, evades autoantibodies in immune-mediated thrombotic thrombocytopenic purpura.

doi: 10.1038/s41598-024-80674-x

Figure Lengend Snippet: Fig. 4. Autoantibody escaping ability of GC1126A using the iTTP patient’s plasma. (a) Relative residual activity of ADAMTS13 and GC1126A in the presence of autoantibodies from iTTP patient’s plasma (n = 2). Each patient’s plasma was diluted to inhibitory antibody concentrations of 0.5, 1, 2, and 3 BU/ml. Each diluted plasma was mixed with the same molar concentration (3 nM) of ADAMTS13 or GC1126A. Relative activity is the ratio of activity remaining after neutralizing antibodies in the patient’s plasma to activity when each substance is free of neutralizing antibodies. (b) Binding level of autoantibodies from patients with iTTP to ADAMTS13 and GC1126A. Statistical significance was determined using the one-way analysis of variance followed by Tukey’s test (α = 0.05). Each bar represents the mean and standard error of the mean (n = 3).

Article Snippet: The recombinant human ADAMTS13 (full-length) protein (rhADAMTS13 FL; R&D Systems, Minnesota, USA) served as the antigen.

Techniques: Clinical Proteomics, Activity Assay, Concentration Assay, Binding Assay

Journal: Scientific reports

Article Title: GC1126A, a novel ADAMTS13 mutein, evades autoantibodies in immune-mediated thrombotic thrombocytopenic purpura.

doi: 10.1038/s41598-024-80674-x

Figure Lengend Snippet: Fig. 5. Relationship between the anti-ADAMTS13 inhibitor titer and GC1126A EC50. The distribution of the amount of GC1126A required to achieve 0.5 IU/ml activity (EC50) for different patient samples at inhibitor titers of 0.6–0.9 BU/ml (n = 4), 1 BU/ml (n = 23), 3 BU/ml (n = 15), 6 BU/ml (n = 9), and 9 BU/ml (n = 5) (the observed data are shown with closed circles). The estimated regression line and the 95% confidence interval are depicted in solid and dotted lines, respectively. The linear regression analysis yielded the following relationship: EC50 (µg/mL) = 0.0347 + 0.0396 × anti-ADAMTS13 inhibitor titer (BU/mL) (R2 = 0.5842).

Article Snippet: The recombinant human ADAMTS13 (full-length) protein (rhADAMTS13 FL; R&D Systems, Minnesota, USA) served as the antigen.

Techniques: Activity Assay

Journal: Scientific reports

Article Title: GC1126A, a novel ADAMTS13 mutein, evades autoantibodies in immune-mediated thrombotic thrombocytopenic purpura.

doi: 10.1038/s41598-024-80674-x

Figure Lengend Snippet: Fig. 6. Comparison of therapeutic efficacy of GC1126A, rh WT-ADAMTS13, and caplacizumab in mitigating platelet count reduction in the iTTP-mimic mouse model. (a) Study design for therapeutic efficacy comparison. (b) Platelet count results. (c) Residual activity of ADAMTS13 in the plasma of an iTTP-mimic mouse model. Values below the limit (0.03 IU/mL) are also not present on the graph. (d) Inhibitory antibody levels (BU levels). Values below the limit (0.42 BU/mL) are also not present on the graph. (b)–(d) Outliers were identified using the Grubbs’ test (α = 0.05) and excluded from each group. Each bar represents the mean and standard error of the mean (n = 4 mice per group).

Article Snippet: The recombinant human ADAMTS13 (full-length) protein (rhADAMTS13 FL; R&D Systems, Minnesota, USA) served as the antigen.

Techniques: Comparison, Drug discovery, Activity Assay, Clinical Proteomics

Journal: Scientific reports

Article Title: GC1126A, a novel ADAMTS13 mutein, evades autoantibodies in immune-mediated thrombotic thrombocytopenic purpura.

doi: 10.1038/s41598-024-80674-x

Figure Lengend Snippet: Fig. 7. Comparison of therapeutic efficacy of GC1126A, rh WT-ADAMTS13, and caplacizumab in restoring severely reduced platelet counts in the iTTP-mimic mouse model. (a) Study design for therapeutic efficacy comparison. (b) Platelet count results. Outliers were identified using the Grubbs’ test (α = 0.05) and excluded from each group. Each bar represents the mean and standard error of the mean (n = 3–10 mice per group).

Article Snippet: The recombinant human ADAMTS13 (full-length) protein (rhADAMTS13 FL; R&D Systems, Minnesota, USA) served as the antigen.

Techniques: Comparison, Drug discovery