human sost protein  (MedChemExpress)

Journal: Frontiers in Immunology

Article Title: In silico design of novel precision vaccine targeting sclerostin epitopes for osteoporosis prevention and treatment

doi: 10.3389/fimmu.2025.1644437

Figure Lengend Snippet: Screening and analysis of high-affinity epitopes on SOST. (A) ELISA experiments were conducted to identify SOST fragments with strong binding affinity for ROMO, revealing that SOST 114–143 and SOST 143–173 exhibit significantly higher affinity ( P <0.01). (B) A schematic diagram delineating the binding functional regions associated with the high-affinity fragments of SOST. (C) ELISA results indicate that SOST 131–163 displays the highest affinity for ROMO ( P <0.01), thereby identifying it as a potent functional epitope of SOST. (D-a) SOST 131–163 fragment (highlighted in yellow) is located within the loop3 domain of SOST protein. (D-b) Docking studies indicate that SOST 131–163 fragment interacts with ROMO light chain, yielding a binding free energy of -25.8 kcal/mol and an interface area of 712.9 Ų. (D-c) Additionally, SOST 131–163 fragment can bind to the ROMO heavy chain, resulting in a binding free energy of -33.19 kcal/mol and an interface area of 451.6 Ų. (E) CTL epitopes within SOST 131–163 sequence include two strong binder epitopes and four weak binder epitopes. (F) HTL epitopes in SOST 131–163 sequence comprise one strong binder epitope and four weak binder epitopes. Predictions of B cell epitopes for SOST 131–163 sequence are illustrated, including predicted linear B cell epitopes (G) and predicted discontinuous B cell epitopes (H) .

Article Snippet: In brief, 1 μg/mL of human SOST protein (MedChemExpress Inc.) was coated onto the wells of MaxiSorp microtiter plates (Thermo Fisher Scientific Inc.) and incubated overnight at 4°C.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Functional Assay, Sequencing

Journal: International journal of molecular sciences

Article Title: The Effect of Sclerostin and Monoclonal Sclerostin Antibody Romosozumab on Osteogenesis and Osteoclastogenesis Mediated by Periodontal Ligament Fibroblasts.

doi: 10.3390/ijms24087574

Figure Lengend Snippet: Figure 2. TRAcP-positive multinucleated cells in HD culture. (A,B) Effect of increasing concentra- tions of sclerostin and romosozumab in a PBMC high density (HD) culture. (A) The number of multinucleated cells formed in a PBMC HD culture in the presence of sclerostin (50–150 ng/mL). The number of multinucleated cells formed in a PBMC HD culture in the presence of romosozumab (0.1–10 µg/mL). No significant differences were observed. PBMCs HD culture experiments were performed in quadruplicate. Mean results ± SEM are presented. Multinucleated cells with more than 3 nuclei were counted and categorized: 3–5, 6–10, and >10 nuclei. Nearly all of the multinucleated cells had 3–5 nuclei; therefore, categories were merged in this figure.

Article Snippet: Recombinant human sclerostin was acquired from R&D Systems (Minneapolis, MN, USA) and dissolved in water.

Techniques:

Journal: International journal of molecular sciences

Article Title: The Effect of Sclerostin and Monoclonal Sclerostin Antibody Romosozumab on Osteogenesis and Osteoclastogenesis Mediated by Periodontal Ligament Fibroblasts.

doi: 10.3390/ijms24087574

Figure Lengend Snippet: Figure 3. Alkaline phosphatase at t = 0 and t = 14 days. Alkaline Phosphatase (ALP) (A,B); DNA (C,D); Alkaline Phosphatase Activity (ALP/DNA) (E,F). (A,B) No statistical differences were observed in ALP between the different concentrations of both sclerostin and romosozumab. (C) Average DNA with increasing concentrations of sclerostin. (D) Average DNA with increasing concentrations of romosozumab. (E) ALP/DNA is slightly, but not significantly, increased with higher concentrations of sclerostin. (F) ALP/DNA appears to decrease slowly. No significant differences were observed in ALP/DNA between the different concentrations of romosozumab. Control c−conditions (days 0 and 14) were created without (−) mineralization medium and with mineralization medium (+) and with different concentrations of sclerostin (0+ to 150+) and romosozumab (0+ to 10+). Assays were performed using three (sclerostin) or five (romosozumab) different donors in duplicates. Average results ± SEM are shown.

Article Snippet: Recombinant human sclerostin was acquired from R&D Systems (Minneapolis, MN, USA) and dissolved in water.

Techniques: Activity Assay, Control

Journal: International journal of molecular sciences

Article Title: The Effect of Sclerostin and Monoclonal Sclerostin Antibody Romosozumab on Osteogenesis and Osteoclastogenesis Mediated by Periodontal Ligament Fibroblasts.

doi: 10.3390/ijms24087574

Figure Lengend Snippet: Figure 4. Alizarin red staining after sclerostin or romosozumab treatment. (A) PDL fibroblasts were cultured without (c−) or with (0+) mineralization medium under different concentrations of sclerostin (ng/mL) at t = 21 (A) or romosozumab at t = 14 (B) and t = 21 (C). Strong heterogeneity was observed between the donors. (D,E) Micrograph of stained PDL fibroblasts. Formation of red nodules is visible under the microscope: donor 1 with addition of 50 ng/mL sclerostin (D); donor 1 with addition of 100 ng/mL sclerostin (E). Red nodules with black speckles indicate mineral deposition. Scale bar represents 100 µm. *: empty wells.

Article Snippet: Recombinant human sclerostin was acquired from R&D Systems (Minneapolis, MN, USA) and dissolved in water.

Techniques: Staining, Cell Culture, Microscopy

Journal: International journal of molecular sciences

Article Title: The Effect of Sclerostin and Monoclonal Sclerostin Antibody Romosozumab on Osteogenesis and Osteoclastogenesis Mediated by Periodontal Ligament Fibroblasts.

doi: 10.3390/ijms24087574

Figure Lengend Snippet: Figure 5. Relative expression of osteogenic markers in a PDL culture without (−) or with (+) min- eralization medium under increasing concentrations of romosozumab. Cells were cultured for 14 days in the absence or presence of romosozumab at 0.1, 1.0, and 10 µg/mL. An amount of 10 µg/mL HI romosozumab was added to the control group with mineralization (0+). RNA was extracted and qPCR was performed to detect the relative expression of (A) RunX2, (B) Col1, (C) ALP, (D) Osteonectin, (E) SOST, and (F) DMP1. (A) RUNX2 shows no significant differences between differ- ent conditions. (B) Col1 shows a significant difference to untreated control (t = 14) for romosozumab at 10 µg/mL. (C) ALP was significantly elevated at t = 14 in the control cultures with and without mineralization in comparison to t = 0. (D) Osteonectin reduced significantly in the 0.1 µm/mL group compared to the control group. (E) SOST showed no significant differences between different condi- tions. (F) Relative expression of DMP1 was comparable between all conditions. Mean results ± SEM of 5 donors are presented. * p < 0.05.

Article Snippet: Recombinant human sclerostin was acquired from R&D Systems (Minneapolis, MN, USA) and dissolved in water.

Techniques: Expressing, Cell Culture, Control, Comparison

Journal: International journal of molecular sciences

Article Title: The Effect of Sclerostin and Monoclonal Sclerostin Antibody Romosozumab on Osteogenesis and Osteoclastogenesis Mediated by Periodontal Ligament Fibroblasts.

doi: 10.3390/ijms24087574

Figure Lengend Snippet: Figure 6. Relative expression of sclerostin and sclerostin receptors LRP 4, 5, and 6 in PDL fibroblasts (n = 5) and in osteocytes (n = 4) from bone isolates. (A) Sclerostin or SOST mRNA expression was significantly higher than that of PDL fibroblasts and 45,000 times the expression of osteocytes. The relative expression of (B) LRP 4 (450×) and (C) LRP 5 (17×) in PDL fibroblasts was significantly lower in comparison to the relative expression of LRP 4 and 5 in osteocytes. (D) Relative mRNA expression of LRP 6 in PDL fibroblasts and osteocytes was comparable. Mean results ± SEM are presented. Significant results are shown with a black bar (* p < 0.05). A Mann–Whitney U test was used.

Article Snippet: Recombinant human sclerostin was acquired from R&D Systems (Minneapolis, MN, USA) and dissolved in water.

Techniques: Expressing, Comparison, MANN-WHITNEY