recombinant human cd40 ligand  (MedChemExpress)

Journal: Nature Communications

Article Title: A bispecific CD40 agonistic antibody allowing for antibody-peptide conjugate formation to enable cancer-specific peptide delivery, resulting in improved T proliferation and anti-tumor immunity in mice

doi: 10.1038/s41467-024-53839-5

Figure Lengend Snippet: A The agonistic activity of the anti-CD40 mAbs clones incubated for 48 h with immature moDCs evaluated for upregulation of markers MHC-II, CD86, and CD83 clustered in heatmap together with IL-12 secretion. B Epitope mapping of the top agonistic clone A9 and non-agonistic clone B1 determined by HDX-MS illustrated in a CD40L-CD40 crystal structure (PDB 3QD6). C Agonistic activity of the mAbs was performed by incubation with immature moDCs for 48 h and evaluated by flow cytometry of CD86 MFI levels (illustrated as background-subtracted log-transformed values) n = 3 (independent donors) and IL-12p40 levels n = 5 (independent donors pooled from two individual experiments) in the supernatant were quantified by ELISA. P -values are shown in the graph (ns = non-significant) and calculated with Kruskal-Wallis with Dunn´s multiple (CD86) and one-way ANOVA with Dunnett´s multiple comparisons test (hIL12p40). D Agonistic activity of mAbs evaluated by upregulation of CD86 on isolated CD19 + B cells via flow cytometry 24 h post-stimulation. n = 2 (independent donors, illustrated as the mean of two technical replicates). E Immature htgCD40 BMDCs were stimulated for 48 h with the mAbs thereafter, IL-12p40 was quantified by ELISA in the supernatant. n = 1 (mean value from two technical replicates). Data is shown as mean ± SEM.

Article Snippet: Human CD40-Fc was diluted to 20 nM in HBS supplemented with 0.05% Tween-20 injected either alone or pre-incubated with 200 nM human CD40L (R&D Systems, Cat. 6420-CLB) and subsequently added.

Techniques: Activity Assay, Clone Assay, Incubation, Flow Cytometry, Transformation Assay, Enzyme-linked Immunosorbent Assay, Isolation