Journal: bioRxiv

Article Title: De novo design of hundreds of functional GPCR-targeting antibodies enabled by scaling test-time compute

doi: 10.1101/2025.05.28.656709

Figure Lengend Snippet: a. On-cell K D measurements for titrations of anti-CXCR4 designs on PathHunter CXCR4 cell line. Error bars are 95% CI for bar plot and SD for binding trace. Weak binders for which an accurate on-cell KD was not calculable are excluded. Binding control is shown in dark grey. b. On-cell K D measurements and titrations of anti-CXCR7 designs on PathHunter CXCR7 cell line as shown in panel (a). c. Binding trace of top anti-CXCR4 design (left) and anti-CXCR7 design (right) against their respective PathHunter target positive (orange) and background (grey) cell line. d . On-cell K D for anti-CXCR4 designs against additional disease-relevant CXCR4 expressing cell lines (Ramos and Jurkat) e. Single-point binding signal for anti-CXCR4 and anti-CXCR7 at their highest expressed concentration against both CXCR4 and CXCR7 cell lines to evaluate on- and off-target binding. f. Developability heatmaps for CXCR4-targeting (top) and CXCR7-targeting (bottom) designs, showing production yield in ExpiCHO post 1-step purification, monomericity by SEC, and polyspecificity by BVP ELISA normalized to Ustekinumab. Designs passing the criteria are denoted by a brown dot. Designs meeting all developability criteria and an on-cell K D threshold of <50 nM are highlighted by an orange box.

Article Snippet: BVP scores were determined by normalizing raw absorbance to control wells with no test antibody (i.e. BVP-only control), and normalized to Trastuzumab (ICH-4031; Ichor Bio) or Ustekinumab (HY-P9909; MedChem Express) scores assayed concurrently.

Techniques: Binding Assay, Control, Expressing, Concentration Assay, Purification, Enzyme-linked Immunosorbent Assay

Journal: European journal of immunology

Article Title: Human rhinoviruses induce IL-35-producing Treg via induction of B7-H1 (CD274) and sialoadhesin (CD169) on DC.

doi: 10.1002/eji.200939527

Figure Lengend Snippet: Figure 4. R-DC induce IL-35 expression, production, and release in human T cells, (A) T cells were cocultured with R-DC (black bars) or DC (white bars) in a 10:1 ratio for 24 h and the production of EBI3, p35, and p28 was measured at the mRNA level via real-time PCR. Data show mean7SD (n 5 2) and are representative of three independent experiments. (B) EBI3 and p35 production in peripheral blood (PB) T cells or naı¨ve T cells isolated from human CB cocultured with DC (open histogram) or R-DC (filled histogram) for 24 h was detected by intracellular staining. The dotted line shows the isotype control mAb VIAP. The T cells gate is shown. Data are representative of three independent experiments. (C) Cell culture SN of T cells stimulated with DC or R-DC for 24–72 h were pooled from six independent experiments and immunoprecipitated with an Ab against p35. These samples were immunoblotted with an EBI3 Ab (upper blot) or a control Ab: lower blot). As a loading control, the light chain of the p35 mAb used for precipitation, detected with sheep-anti-mouse-HRP (SaM), was used. Blots were further analyzed densiometrically using Image J software. Data are presented as ratio of EBI3 protein to light chain. This experiment was performed three times and is representative of a pool of six cell donors. (D) As in (C) but the anti-p35 coated beads left after precipitation were analyzed via flow cytometry. IL-35 was detected with an anti-EBI3 Ab. (E) As in (D) but the IL-35 loaded anti-p35 coated beads were stained with anti-IL-12p40 and anti-IL-27 mAb. This experiment was performed once and is representative of a pool of six cell donors; po0.01 and po0.001 (D and E).

Article Snippet: The monoclonal Ab against p35 (clone27537), IL-12p40 (mAb609), IL-12p70 (mAb611), IL-27 (mAb25261), and TGF-b (mAb240) were purchased from R&D Systems.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Isolation, Staining, Control, Cell Culture, Immunoprecipitation, Software, Cytometry