sost protein  (Bio-Techne corporation)

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: bioRxiv

Article Title: Spatial transcriptomics for gene discovery identifies Slc13a5 as a modulator of bone mechanoadaptation

doi: 10.64898/2026.03.11.711126

Figure Lengend Snippet: Relative expression of Slc13a5 in primary osteoblasts culture treated with recombinant sclerostin (SOST) or PBS vehicle (Veh). Graph represents mean±SD; Student’s t-test versus control, ***p<0.001, (n=3).

Article Snippet: For sclerostin treatment, differentiated pCOBs were serum-reduced to 0.1% FBS and treated with 100ng/ml of recombinant mouse sclerostin (R&D systems) or vehicle control (PBS) for 24 h. Total RNA was isolated from cultured cells following lysis in RLT buffer (Qiagen) using the RNeasy Micro Kit (Qiagen) in accordance with the manufacturer’s guidelines.

Techniques: Expressing, Recombinant, Control

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: During the second medium change, anti-SOST antiserum and 100 ng/mL recombinant SOST protein (Novoprotein) were added.

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

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: Construction plan and immune stimulation simulation of SOST candidate vaccines. (A) Schematic representation for the construction of SOST candidate vaccines. (B) Predicted three-dimensional structure of SOST candidate vaccines, modeled using AlphaFold2 server based on amino acid sequence. The cyan region denotes DTT protein scaffold, while the yellow regions represent the various repeated SOST 131–163 peptides. (C) Immune stimulation simulation conducted using C-IMMSIM online server demonstrates that DS 3 and DS 5 vaccines simultaneously stimulate the production of IgM, IgG1, and IgG2 (Romosozumab is classified as an IgG2 antibody), whereas other candidate vaccines primarily induced IgM and IgG1 antibodies. Consequently, DS 3 and DS 5 vaccines were selected for further analysis.

Article Snippet: During the second medium change, anti-SOST antiserum and 100 ng/mL recombinant SOST protein (Novoprotein) were added.

Techniques: Vaccines, Sequencing

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: Cloning, expression, and immunogenicity assessment of DS 3 and DS 5 vaccines. (A) DS 3 vaccine sequence (red) was inserted into the pSmartI expression vector (black) via seamless cloning using XhoI. (B) Agarose gel electrophoresis showing: Lane 1, recombinant plasmid; Lane 2, target fragment (1268 bp) along with vector sequence; Lane M, 1 kb DNA ladder. (C) Expression and purification of DS 3 vaccine. (D) DS 5 vaccine sequence (red) was cloned into the pSmartI expression vector (black) via seamless cloning using XhoI. (E) Agarose gel electrophoresis illustrating: Lane 1, recombinant plasmid; Lane 2, target fragment (1520 bp) along with vector sequence; Lane M, 1 kb DNA ladder. (F) Expression and purification of DS 5 . (G) Schematic overview of the mouse immunization protocol; each group comprised three mice (n=3), serving as independent biological replicates. (H) ELISA measurements indicating significantly elevated serum titers of anti-SOST antibodies in mice immunized with DS 3 and DS 5 compared to PBS controls (serum dilution 1:200). Antibody assays were performed in technical duplicates per mouse. (I–K) Cytokine levels of IL-4, IL-10, and IFN-γ in supernatants from splenocyte stimulation assays. Data are expressed as mean ± SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test. ( * p < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns = no significance).

Article Snippet: During the second medium change, anti-SOST antiserum and 100 ng/mL recombinant SOST protein (Novoprotein) were added.

Techniques: Cloning, Expressing, Immunopeptidomics, Vaccines, Sequencing, Plasmid Preparation, Agarose Gel Electrophoresis, Recombinant, Purification, Clone Assay, Enzyme-linked Immunosorbent Assay

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: Effects of anti-SOST antiserum derived from vaccine-immunized mice on osteoclast and osteoblast differentiation. (A) TRAP staining of bone marrow-derived macrophages treated with SOST and anti-SOST antiserum at a 1:500 dilution, demonstrating inhibition of osteoclast differentiation (n=4). (B) Quantification of TRAP-positive osteoclasts in (A) . (C) Alizarin Red S staining of bone marrow mesenchymal stem cell-derived osteoblasts cultured with osteogenic medium and treated with SOST and anti-SOST antiserum at a 1:100 dilution, indicating enhanced osteoblast differentiation and mineralization upon vaccine antiserum treatment (n=4). (D) Quantitative analysis of mineralization in (C) . (E) Alizarin Red S staining of MC3T3-E1 subclone 14 cells cultured with osteogenic medium and treated with SOST and vaccine antiserum at a 1:100 dilution, showing rescue of SOST-mediated inhibition of osteoblast differentiation and mineralization (n=6). (F) Quantitative analysis of mineralization in (E) . Scale bars, 250 μm and 500 μm. Data are expressed as mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test.

Article Snippet: During the second medium change, anti-SOST antiserum and 100 ng/mL recombinant SOST protein (Novoprotein) were added.

Techniques: Derivative Assay, Staining, Inhibition, Cell Culture