Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Characterization of VHH clone diversity. ( A , B ) Dendrograms resulting from multiple sequence alignment of amino acid sequences of VHH clones specific for IL-6 ( A ) and IL-17A ( B ), performed using Clustal Omega [ https://www.ebi.ac.uk/jdispatcher/msa/clustalo ] (EMBL-EBI service providing bioinformatics tools; Accessed on 22 January 2026) . An asterisk (*) indicates clones that, at the phage library stage, contained a STOP codon in the coding sequence and required the presence of a corresponding suppressor tRNA provided by the E. coli host cell for protein synthesis. In red, two clones are marked that were selected at the end of the development stage for final BsAb assembly. ( C , D ) Residue conservation presented as a sequence logo representation of complementarity-determining regions’ (CDRs) repertoire among anti-IL-6 clones ( C ) and anti-IL-17A ones ( D ) obtained using WebLogo 3, a web-based application [ https://weblogo.threeplusone.com/create.cgi ] (accessed on 22 January 2026) [ , ]. The overall height of the stack at each position indicates the sequence conservation at that position, while the height of amino acid symbols within the stack represents the relative frequency of each amino acid. Moreover, the width of the stack is proportional to the fraction of the amino acid presence at a particular position, giving a richer picture of sequence conservation. The color scheme used shows amino acids according to their chemical properties.

Article Snippet: HEK-Blue IL-6 (#hkb-il6; InvivoGen, San Diego, CA, USA) or HEK-Blue IL-17 (#hkb-il17; InvivoGen) reporter cells were used to assess the biological activity of VHHs.

Techniques: Sequencing, Clone Assay, Residue

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Dissociation rates (k off in 1/s) of VHH clones specific for anti-IL-17A ( A ) and anti-IL-6 ( B ), were analyzed using bio-layer interferometry (BLI). The data illustrate the variability in dissociations rate across the selected clones for both IL-17A and IL-6. The clones highlighted in red, D3 for IL-17A and A5 for IL-6, represent the lead clones selected at a later stage of development.

Article Snippet: HEK-Blue IL-6 (#hkb-il6; InvivoGen, San Diego, CA, USA) or HEK-Blue IL-17 (#hkb-il17; InvivoGen) reporter cells were used to assess the biological activity of VHHs.

Techniques: Clone Assay

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: SDS-PAGE analysis of VHH clones under reducing conditions after purification using the KingFisher Flex station. Protein samples were loaded at 1 µg per well. Representative SDS-PAGE profiles of anti-IL-17A clones ( A ) and anti-IL-6 clones ( B ) illustrate the purity and integrity of the purified clones. The most promising VHHs, D3 for IL-17A and A5 for IL-6, are marked in red.

Article Snippet: HEK-Blue IL-6 (#hkb-il6; InvivoGen, San Diego, CA, USA) or HEK-Blue IL-17 (#hkb-il17; InvivoGen) reporter cells were used to assess the biological activity of VHHs.

Techniques: SDS Page, Clone Assay, Purification

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Purity analysis for all purified candidates from among anti-IL-17A clones ( A ) and anti-IL-6 clones ( B ) using capillary electrophoresis (LabChip system). The most promising VHHs, D3 for IL-17A and A5 for IL-6, are marked in red. Exemplary overlapped data analysis results obtained from the capillary electrophoresis are shown for anti-IL-17A clones ( C ) and anti-IL-6 clones ( D ).

Article Snippet: HEK-Blue IL-6 (#hkb-il6; InvivoGen, San Diego, CA, USA) or HEK-Blue IL-17 (#hkb-il17; InvivoGen) reporter cells were used to assess the biological activity of VHHs.

Techniques: Purification, Clone Assay, Electrophoresis

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Melting temperature determination ( left panels ) for anti-IL-17A ( A ) and anti-IL-6 clones ( B ) performed using the Uncle platform, which enables the assessment of the thermal stability of the purified candidates. The most promising VHHs, D3 for IL-17A and A5 for IL-6, are marked in red. Representative melting curve analysis results for selected anti-IL-17A and anti-IL-6 clones are presented on the right panels .

Article Snippet: HEK-Blue IL-6 (#hkb-il6; InvivoGen, San Diego, CA, USA) or HEK-Blue IL-17 (#hkb-il17; InvivoGen) reporter cells were used to assess the biological activity of VHHs.

Techniques: Clone Assay, Purification

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: The range of KD values for VHH clones specific to anti-IL-17A ( A ) and anti-IL-6 ( B ). The most promising VHHs, D3 for IL-17A and A5 for IL-6, are marked in red on the left panel . On the right panel , representative examples of BLI data analysis are shown. The red lines indicate the fitted curves, representing the global fitting of the binding data to a 1:1 interaction model.

Article Snippet: HEK-Blue IL-6 (#hkb-il6; InvivoGen, San Diego, CA, USA) or HEK-Blue IL-17 (#hkb-il17; InvivoGen) reporter cells were used to assess the biological activity of VHHs.

Techniques: Clone Assay, Binding Assay

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Neutralization potency of selected VHH clones targeting IL-17A ( A ) and IL-6 ( B ). The range of IC 50 values of VHHs is shown on the left panel . The most promising VHHs, D3 for IL-17A and A5 for IL-6, are marked in red. Right panel shows representative IC 50 determination analyses using reporter cell assays, where the dose-dependent inhibition of cytokine signaling was measured. Data were analyzed using nonlinear regression with a three-parameter dose–response model (log [inhibitor] vs. response), fitted using GraphPad Prism software. Each experiment was performed in two technical replicates.

Article Snippet: HEK-Blue IL-6 (#hkb-il6; InvivoGen, San Diego, CA, USA) or HEK-Blue IL-17 (#hkb-il17; InvivoGen) reporter cells were used to assess the biological activity of VHHs.

Techniques: Neutralization, Clone Assay, Inhibition, Software

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Relationship between IC 50 values from in vitro assays and binding affinity measured by BLI. ( A ) Correlation for IL-17A, with a correlation coefficient (R 2 ) of 0.8297. ( B ) Similar strong correlation for IL-6, as indicated by an R 2 of 0.7714. The most promising VHHs, D3 for IL-17A and A5 for IL-6, are marked in red.

Article Snippet: HEK-Blue IL-6 (#hkb-il6; InvivoGen, San Diego, CA, USA) or HEK-Blue IL-17 (#hkb-il17; InvivoGen) reporter cells were used to assess the biological activity of VHHs.

Techniques: In Vitro, Binding Assay

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Binding affinity analysis of the CPBT0853 bispecific antibody. Representative bio-layer interferometry (BLI) binding curves for CPBT0853 interacting with human IL-17A ( A ), human IL-6 ( B ), human IL-17A/F heterodimer ( C ), cynomolgus monkey IL-17A ( D ), and cynomolgus monkey IL-6 ( E ) are shown. The absence of binding is demonstrated by the lack of interaction between CPBT0853 and mouse IL-6 ( F ).

Article Snippet: HEK-Blue IL-6 (#hkb-il6; InvivoGen, San Diego, CA, USA) or HEK-Blue IL-17 (#hkb-il17; InvivoGen) reporter cells were used to assess the biological activity of VHHs.

Techniques: Binding Assay

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Blocking activity of the bispecific antibody CPBT0853 measured by bio-layer interferometry (BLI). Bar graphs represent quantitative data derived from BLI sensorgrams showing the effect of increasing concentrations of CPBT0853 on the binding of IL-17A ( A ) or IL-6 ( B ) to their respective receptors. The measured response [nm], reflecting the amount of receptor bound to the immobilized cytokine, decreases with increasing antibody concentration, indicating dose-dependent inhibition of cytokine–receptor interactions. Data represent mean ± SD from three independent experiments.

Article Snippet: HEK-Blue IL-6 (#hkb-il6; InvivoGen, San Diego, CA, USA) or HEK-Blue IL-17 (#hkb-il17; InvivoGen) reporter cells were used to assess the biological activity of VHHs.

Techniques: Blocking Assay, Activity Assay, Derivative Assay, Binding Assay, Concentration Assay, Inhibition

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Neutralization potency of CPBT0853 targeting IL-17A ( A ) and IL-6, IL-17A. Representative graphs show IC 50 analysis performed using a reporter cell assay. For comparison, parental VHHs and monospecific referential antibodies were used. The cellular response, measured as cytokine-induced activation, progressively declined with increasing concentrations of the tested antibodies, indicating effective inhibition of signaling through IL-6 ( A ), IL-17A ( B ), and the IL-17A/F heterodimer ( C ). Data were analyzed using nonlinear regression with a three-parameter dose–response model (log [inhibitor] vs. response), fitted using GraphPad Prism software. The curves shown represent a representative biological replicate performed with two technical replicates. The IC 50 values summarized in were calculated from three independent biological replicates.

Article Snippet: HEK-Blue IL-6 (#hkb-il6; InvivoGen, San Diego, CA, USA) or HEK-Blue IL-17 (#hkb-il17; InvivoGen) reporter cells were used to assess the biological activity of VHHs.

Techniques: Neutralization, Comparison, Activation Assay, Inhibition, Software

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Representative IC 50 curves showing inhibition of STATs phosphorylation in PBMCs stimulated with IL-6 and treated with increasing concentrations of the bispecific antibody CPBT0853. Phosphorylation of STAT1 ( A ) and STAT3 ( B ) was measured by flow cytometry. Data were normalized to cytokine-only controls and expressed as % pSTAT + lymphocytes. IC 50 values were calculated using a nonlinear regression with a three-parameter dose–response model (log [inhibitor] vs. response) in GraphPad Prism software. Each curve represents technical duplicates from PBMCs of one representative donor out of three tested.

Article Snippet: HEK-Blue IL-6 (#hkb-il6; InvivoGen, San Diego, CA, USA) or HEK-Blue IL-17 (#hkb-il17; InvivoGen) reporter cells were used to assess the biological activity of VHHs.

Techniques: Inhibition, Phospho-proteomics, Flow Cytometry, Software

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Inhibition of cytokine production (IL-6 and IL-8) in HFLS cells by CPBT0853. Levels of IL-6 ( A ) and IL-8 ( B ) secreted into the culture medium after stimulation of the cells with IL-17A. Additionally, cells were treated with bispecific and monospecific antibodies, as indicated. Data are shown as the mean ± SD from a minimum of four independent experiments. The bispecific antibody CPBT0853 effectively reduced the IL-6 and IL-8 levels, with higher efficiency, as compared to the monospecific antibodies, indicating enhanced anti-inflammatory activity. Additionally, the humanized IgG4 form of CPBT0853 (CPBT1269) was evaluated for comparison, showing activity in cytokine inhibition similar to that of the parental/original IgG1 form.

Article Snippet: HEK-Blue IL-6 (#hkb-il6; InvivoGen, San Diego, CA, USA) or HEK-Blue IL-17 (#hkb-il17; InvivoGen) reporter cells were used to assess the biological activity of VHHs.

Techniques: Inhibition, Activity Assay, Comparison

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Characterization of VHH clone diversity. ( A , B ) Dendrograms resulting from multiple sequence alignment of amino acid sequences of VHH clones specific for IL-6 ( A ) and IL-17A ( B ), performed using Clustal Omega [ https://www.ebi.ac.uk/jdispatcher/msa/clustalo ] (EMBL-EBI service providing bioinformatics tools; Accessed on 22 January 2026) . An asterisk (*) indicates clones that, at the phage library stage, contained a STOP codon in the coding sequence and required the presence of a corresponding suppressor tRNA provided by the E. coli host cell for protein synthesis. In red, two clones are marked that were selected at the end of the development stage for final BsAb assembly. ( C , D ) Residue conservation presented as a sequence logo representation of complementarity-determining regions’ (CDRs) repertoire among anti-IL-6 clones ( C ) and anti-IL-17A ones ( D ) obtained using WebLogo 3, a web-based application [ https://weblogo.threeplusone.com/create.cgi ] (accessed on 22 January 2026) [ , ]. The overall height of the stack at each position indicates the sequence conservation at that position, while the height of amino acid symbols within the stack represents the relative frequency of each amino acid. Moreover, the width of the stack is proportional to the fraction of the amino acid presence at a particular position, giving a richer picture of sequence conservation. The color scheme used shows amino acids according to their chemical properties.

Article Snippet: HEK-Blue IL-6 reporter cells (#hkb-il6; InvivoGen) or HEK-Blue IL-17 reporter cells (#hkb-il17; InvivoGen) were used to assess the activity of the bispecific antibody CPBT0853, respectively.

Techniques: Sequencing, Clone Assay, Residue

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Dissociation rates (k off in 1/s) of VHH clones specific for anti-IL-17A ( A ) and anti-IL-6 ( B ), were analyzed using bio-layer interferometry (BLI). The data illustrate the variability in dissociations rate across the selected clones for both IL-17A and IL-6. The clones highlighted in red, D3 for IL-17A and A5 for IL-6, represent the lead clones selected at a later stage of development.

Article Snippet: HEK-Blue IL-6 reporter cells (#hkb-il6; InvivoGen) or HEK-Blue IL-17 reporter cells (#hkb-il17; InvivoGen) were used to assess the activity of the bispecific antibody CPBT0853, respectively.

Techniques: Clone Assay

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: SDS-PAGE analysis of VHH clones under reducing conditions after purification using the KingFisher Flex station. Protein samples were loaded at 1 µg per well. Representative SDS-PAGE profiles of anti-IL-17A clones ( A ) and anti-IL-6 clones ( B ) illustrate the purity and integrity of the purified clones. The most promising VHHs, D3 for IL-17A and A5 for IL-6, are marked in red.

Article Snippet: HEK-Blue IL-6 reporter cells (#hkb-il6; InvivoGen) or HEK-Blue IL-17 reporter cells (#hkb-il17; InvivoGen) were used to assess the activity of the bispecific antibody CPBT0853, respectively.

Techniques: SDS Page, Clone Assay, Purification

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Purity analysis for all purified candidates from among anti-IL-17A clones ( A ) and anti-IL-6 clones ( B ) using capillary electrophoresis (LabChip system). The most promising VHHs, D3 for IL-17A and A5 for IL-6, are marked in red. Exemplary overlapped data analysis results obtained from the capillary electrophoresis are shown for anti-IL-17A clones ( C ) and anti-IL-6 clones ( D ).

Article Snippet: HEK-Blue IL-6 reporter cells (#hkb-il6; InvivoGen) or HEK-Blue IL-17 reporter cells (#hkb-il17; InvivoGen) were used to assess the activity of the bispecific antibody CPBT0853, respectively.

Techniques: Purification, Clone Assay, Electrophoresis

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Melting temperature determination ( left panels ) for anti-IL-17A ( A ) and anti-IL-6 clones ( B ) performed using the Uncle platform, which enables the assessment of the thermal stability of the purified candidates. The most promising VHHs, D3 for IL-17A and A5 for IL-6, are marked in red. Representative melting curve analysis results for selected anti-IL-17A and anti-IL-6 clones are presented on the right panels .

Article Snippet: HEK-Blue IL-6 reporter cells (#hkb-il6; InvivoGen) or HEK-Blue IL-17 reporter cells (#hkb-il17; InvivoGen) were used to assess the activity of the bispecific antibody CPBT0853, respectively.

Techniques: Clone Assay, Purification

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: The range of KD values for VHH clones specific to anti-IL-17A ( A ) and anti-IL-6 ( B ). The most promising VHHs, D3 for IL-17A and A5 for IL-6, are marked in red on the left panel . On the right panel , representative examples of BLI data analysis are shown. The red lines indicate the fitted curves, representing the global fitting of the binding data to a 1:1 interaction model.

Article Snippet: HEK-Blue IL-6 reporter cells (#hkb-il6; InvivoGen) or HEK-Blue IL-17 reporter cells (#hkb-il17; InvivoGen) were used to assess the activity of the bispecific antibody CPBT0853, respectively.

Techniques: Clone Assay, Binding Assay

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Neutralization potency of selected VHH clones targeting IL-17A ( A ) and IL-6 ( B ). The range of IC 50 values of VHHs is shown on the left panel . The most promising VHHs, D3 for IL-17A and A5 for IL-6, are marked in red. Right panel shows representative IC 50 determination analyses using reporter cell assays, where the dose-dependent inhibition of cytokine signaling was measured. Data were analyzed using nonlinear regression with a three-parameter dose–response model (log [inhibitor] vs. response), fitted using GraphPad Prism software. Each experiment was performed in two technical replicates.

Article Snippet: HEK-Blue IL-6 reporter cells (#hkb-il6; InvivoGen) or HEK-Blue IL-17 reporter cells (#hkb-il17; InvivoGen) were used to assess the activity of the bispecific antibody CPBT0853, respectively.

Techniques: Neutralization, Clone Assay, Inhibition, Software

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Relationship between IC 50 values from in vitro assays and binding affinity measured by BLI. ( A ) Correlation for IL-17A, with a correlation coefficient (R 2 ) of 0.8297. ( B ) Similar strong correlation for IL-6, as indicated by an R 2 of 0.7714. The most promising VHHs, D3 for IL-17A and A5 for IL-6, are marked in red.

Article Snippet: HEK-Blue IL-6 reporter cells (#hkb-il6; InvivoGen) or HEK-Blue IL-17 reporter cells (#hkb-il17; InvivoGen) were used to assess the activity of the bispecific antibody CPBT0853, respectively.

Techniques: In Vitro, Binding Assay

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Binding affinity analysis of the CPBT0853 bispecific antibody. Representative bio-layer interferometry (BLI) binding curves for CPBT0853 interacting with human IL-17A ( A ), human IL-6 ( B ), human IL-17A/F heterodimer ( C ), cynomolgus monkey IL-17A ( D ), and cynomolgus monkey IL-6 ( E ) are shown. The absence of binding is demonstrated by the lack of interaction between CPBT0853 and mouse IL-6 ( F ).

Article Snippet: HEK-Blue IL-6 reporter cells (#hkb-il6; InvivoGen) or HEK-Blue IL-17 reporter cells (#hkb-il17; InvivoGen) were used to assess the activity of the bispecific antibody CPBT0853, respectively.

Techniques: Binding Assay

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Blocking activity of the bispecific antibody CPBT0853 measured by bio-layer interferometry (BLI). Bar graphs represent quantitative data derived from BLI sensorgrams showing the effect of increasing concentrations of CPBT0853 on the binding of IL-17A ( A ) or IL-6 ( B ) to their respective receptors. The measured response [nm], reflecting the amount of receptor bound to the immobilized cytokine, decreases with increasing antibody concentration, indicating dose-dependent inhibition of cytokine–receptor interactions. Data represent mean ± SD from three independent experiments.

Article Snippet: HEK-Blue IL-6 reporter cells (#hkb-il6; InvivoGen) or HEK-Blue IL-17 reporter cells (#hkb-il17; InvivoGen) were used to assess the activity of the bispecific antibody CPBT0853, respectively.

Techniques: Blocking Assay, Activity Assay, Derivative Assay, Binding Assay, Concentration Assay, Inhibition

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Neutralization potency of CPBT0853 targeting IL-17A ( A ) and IL-6, IL-17A. Representative graphs show IC 50 analysis performed using a reporter cell assay. For comparison, parental VHHs and monospecific referential antibodies were used. The cellular response, measured as cytokine-induced activation, progressively declined with increasing concentrations of the tested antibodies, indicating effective inhibition of signaling through IL-6 ( A ), IL-17A ( B ), and the IL-17A/F heterodimer ( C ). Data were analyzed using nonlinear regression with a three-parameter dose–response model (log [inhibitor] vs. response), fitted using GraphPad Prism software. The curves shown represent a representative biological replicate performed with two technical replicates. The IC 50 values summarized in were calculated from three independent biological replicates.

Article Snippet: HEK-Blue IL-6 reporter cells (#hkb-il6; InvivoGen) or HEK-Blue IL-17 reporter cells (#hkb-il17; InvivoGen) were used to assess the activity of the bispecific antibody CPBT0853, respectively.

Techniques: Neutralization, Comparison, Activation Assay, Inhibition, Software

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Representative IC 50 curves showing inhibition of STATs phosphorylation in PBMCs stimulated with IL-6 and treated with increasing concentrations of the bispecific antibody CPBT0853. Phosphorylation of STAT1 ( A ) and STAT3 ( B ) was measured by flow cytometry. Data were normalized to cytokine-only controls and expressed as % pSTAT + lymphocytes. IC 50 values were calculated using a nonlinear regression with a three-parameter dose–response model (log [inhibitor] vs. response) in GraphPad Prism software. Each curve represents technical duplicates from PBMCs of one representative donor out of three tested.

Article Snippet: HEK-Blue IL-6 reporter cells (#hkb-il6; InvivoGen) or HEK-Blue IL-17 reporter cells (#hkb-il17; InvivoGen) were used to assess the activity of the bispecific antibody CPBT0853, respectively.

Techniques: Inhibition, Phospho-proteomics, Flow Cytometry, Software

Journal: Antibodies

Article Title: Development of Bispecific Antibody Targeting Human IL-17A and IL-6

doi: 10.3390/antib15020029

Figure Lengend Snippet: Inhibition of cytokine production (IL-6 and IL-8) in HFLS cells by CPBT0853. Levels of IL-6 ( A ) and IL-8 ( B ) secreted into the culture medium after stimulation of the cells with IL-17A. Additionally, cells were treated with bispecific and monospecific antibodies, as indicated. Data are shown as the mean ± SD from a minimum of four independent experiments. The bispecific antibody CPBT0853 effectively reduced the IL-6 and IL-8 levels, with higher efficiency, as compared to the monospecific antibodies, indicating enhanced anti-inflammatory activity. Additionally, the humanized IgG4 form of CPBT0853 (CPBT1269) was evaluated for comparison, showing activity in cytokine inhibition similar to that of the parental/original IgG1 form.

Article Snippet: HEK-Blue IL-6 reporter cells (#hkb-il6; InvivoGen) or HEK-Blue IL-17 reporter cells (#hkb-il17; InvivoGen) were used to assess the activity of the bispecific antibody CPBT0853, respectively.

Techniques: Inhibition, Activity Assay, Comparison

Journal: bioRxiv

Article Title: A modular mRNA–LNP vaccine platform enables integrated RNA, lipid and antigen design to protect against CCHFV

doi: 10.64898/2026.01.17.699915

Figure Lengend Snippet: Innate activation was compared across mRNA–LNPs formulated with BP-104, SM-102, or ALC-0315 using NPmut mRNA as a common payload, and across matched empty LNP controls (iLNP; no mRNA) to isolate lipid-driven signaling. Reporter cells were exposed to a dose range of formulations (1 μg, 500 ng, 250 ng, 125 ng, and 62.5 ng; as indicated), and pathway activation was quantified as fold-change induction relative to baseline (dotted line). Bars denote mean responses with error bars indicating variability across replicates; selected pairwise comparisons are annotated with exact P values. a, NF-κB activation measured in THP1-Dual™ cells as induction of NF-κB–SEAP. Across doses, NPmut mRNA/LNPs and empty LNPs produced modest but consistent NF-κB induction, with broadly similar activation profiles across BP-104-, SM-102-, and ALC-0315–containing LNPs. b, IL-6 pathway signaling measured in HEKBlue–IL-6 reporter cells as fold-change induction of IL-6–dependent signaling. All three ionizable lipid formulations elicited comparable IL-6 activation across the dilution series, and empty LNP controls also induced IL-6 signaling, indicating that both lipid composition and mRNA cargo contribute to pathway engagement. c, Type I interferon signaling measured in IFN-α/β reporter HEK293 cells as fold-change induction of IFN-I activity. NPmut mRNA/LNPs triggered IFN-I responses across lipids, with ALC-0315 producing slightly higher induction at select concentrations (P values shown). Empty LNPs also elicited measurable IFN-I signaling, consistent with lipid-dependent innate stimulation independent of mRNA. Collectively, these data show that BP-104 engages canonical innate immune pathways (NF-κB, IL-6, and type I interferon) at levels comparable to SM-102 and ALC-0315 under controlled dosing conditions, supporting its suitability as an ionizable lipid for mRNA–LNP vaccine delivery.

Article Snippet: HEK-BlueTM IFN-α/β (InvivoGen, Cat# hkb-ifnabv2), HEK-BlueTM IL-6 (InvivoGen, Cat# hkb-hil6), and THP1-DualTM (InvivoGen, Cat# thpd-nfis) were employed to assess type I interferon activity and innate immune activation in response to NPmut mRNA–LNPs formulated with BP-104, SM-102, or ALC-0315.

Techniques: Activation Assay, Produced, Activity Assay

Journal: bioRxiv

Article Title: A modular mRNA–LNP vaccine platform enables integrated RNA, lipid and antigen design to protect against CCHFV

doi: 10.64898/2026.01.17.699915

Figure Lengend Snippet: Primary murine dendritic cells (DCs) were loaded with NPmut mRNA formulated in lipid nanoparticles (LNPs) containing BP-104, SM-102, or ALC-0315, then co-cultured with naïve CD4⁺ and CD8⁺ T cells to quantify intracellular cytokines and soluble mediators in supernatants. a, Frequencies of cytokine-positive CD4⁺ and CD8⁺ T cells measured by intracellular cytokine staining, including IFN-γ and TNF-α (top), IL-2⁺ CD8⁺ cells (bottom left), and IL-17A⁺ CD4⁺ cells (bottom right). Bars show group means with individual biological replicates overlaid; statistical comparisons and corresponding P values are indicated. Across formulations, NPmut mRNA/LNP-loaded DCs elicited Th1-skewed responses (IFN-γ and TNF-α) with a measurable IL-17A⁺ CD4⁺ population, whereas negative control cultures remained low. ALC-0315 drove the strongest IFN-γ⁺ and TNF-α⁺ T cell responses across CD4⁺ and CD8⁺ compartments and increased IL-17A⁺ CD4⁺ frequencies, BP-104 produced intermediate activation, and SM-102 was comparatively attenuated. Lipid-dependent differences were also evident for IL-2⁺ CD8⁺ responses, indicating differential programming of proliferative/expansion-associated outputs. b, Secreted cytokine and chemokine profiles in DC–T cell co-culture supernatants (mean fluorescence intensity, MFI) measured for inflammatory and recruitment-associated mediators, including TNF-α, CXCL1, IFN-γ, IL-12, CCL5, IL-4, CXCL10, GM-CSF, IL-10, IL-6, IP-10, and MCP-1. Error bars indicate variability across replicates; selected pairwise comparisons are annotated with P values. Supernatant signatures mirrored intracellular T cell phenotypes: ALC-0315 induced the most pronounced inflammatory/recruitment program, BP-104 generated a coherent but moderated profile, similar to SM-102. Together, these data show that ionizable lipid identity imprints innate cytokine/chemokine cues that scale and shape downstream Th1/Th17 polarization under fixed mRNA payload and DC-loading conditions.

Article Snippet: HEK-BlueTM IFN-α/β (InvivoGen, Cat# hkb-ifnabv2), HEK-BlueTM IL-6 (InvivoGen, Cat# hkb-hil6), and THP1-DualTM (InvivoGen, Cat# thpd-nfis) were employed to assess type I interferon activity and innate immune activation in response to NPmut mRNA–LNPs formulated with BP-104, SM-102, or ALC-0315.

Techniques: Cell Culture, Staining, Negative Control, Produced, Activation Assay, Co-Culture Assay, Fluorescence, Generated