Journal: Journal of Translational Medicine

Article Title: A humanized LIF-targeting antibody synergistically enhances chemo- and immunotherapy in pancreatic cancer

doi: 10.1186/s12967-026-07723-z

Figure Lengend Snippet: Anti-LIF antibodies inhibit pancreatic cancer cell viability by blocking the LIF-induced STAT3 phosphorylation. A . Purity of the humanized anti-LIF antibody, assessed by SDS‒PAGE followed by Coomassie Brilliant Blue staining. B . Binding affinity of the humanized anti-LIF antibody to LIF, as determined by surface plasmon resonance (SPR) using a biacore 8K system with a protein a chip, yielding a dissociation constant (KD) of 4.43 × 10 − 1 2 M, Kon of 3.86 × 10 6 M − 1 s − 1 , and Koff of 1.71 × 10 −7 s − 1 . C . Western blot analysis showing the inhibitory effect of the humanized anti-LIF antibody on STAT3 phosphorylation induced by exogenous LIF protein. D . Western blot analysis showing the inhibitory effect of the humanized anti-LIF antibody on STAT3 phosphorylation induced by endogenous LIF protein, as well as the rescue effect of exogenous LIF protein. E . The humanized anti-LIF antibody exhibits superior anti-proliferative activity in 2D monolayer cultures. Cell viability (assessed via CCK-8 assay) is reduced more effectively by the humanized anti-LIF antibody compared to the murine anti-LIF antibody, with isotype IgG as the control. F . The humanized anti-LIF antibody inhibits viability in 3D spheroid models. Cells seeded in Matrigel and treated with the humanized anti-LIF antibody show greater viability reduction (assessed via CellTiter-Glo 2.0 assay) compared to the murine anti-LIF antibody, with isotype IgG as the control. All the experiments were performed in triplicate ( n = 3). Statistical analysis was conducted using one-way ANOVA with Tukey’s multiple comparison test or games-howell post-hoc comparisons. Significance was indicated as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Antigen-binding affinity was quantified via surface plasmon resonance (SPR) using a Biacore 8K system (Cytiva) with a Series S Sensor Chip Protein A (Cytiva) for antibody immobilization.

Techniques: Blocking Assay, Phospho-proteomics, Staining, Binding Assay, SPR Assay, Western Blot, Activity Assay, CCK-8 Assay, Control, Comparison