Journal: Communications Biology
Article Title: An engineered ligand trap inhibits leukemia inhibitory factor as pancreatic cancer treatment strategy
doi: 10.1038/s42003-021-01928-2
Figure Lengend Snippet: a Yeast-displayed eLIFR containing the CBM I–Ig-like–CBM II domains (blue circles) has a slightly higher affinity for LIF-His than eLIFR CBM I–Ig-like domains (gray squares). Data are the mean ± SD of triplicate measurements. Data from individual experiments are shown as faint symbols. b Schematic of hLIFR-Fc and eLIFR-Fc. The three N-terminal domains, CBM I–Ig-like–CBM II, are fused to an hIgG1 Fc-domain, with the engineered Ig-like domain shown in dark teal for eLIFR-Fc. c Versus hLIFR-Fc, eLIFR-Fc remains more strongly bound to yeast-displayed LIF after overnight incubation with soluble LIF competitor, indicating a slower off-rate. * P = 0.024, ** P = 0.0024 versus the corresponding hLIFR-Fc condition, two-tailed unpaired Student’s t test. Data are mean ± SD ( n = 3). d KinExA data showing that recombinant eLIFR-Fc (blue triangles) has higher affinity to soluble hLIF-His than hLIFR-Fc (brown circles). Data are the mean of duplicate measurements. Data from individual experiments are shown as faint symbols. e hLIFR-Fc, eLIFR-Fc (bivalent and one-arm), and D25 antibody compete LIF away from WT LIFR. ns not significant, P = 0.03 for one-arm eLIFR-Fc and P = 0.04 for eLIFR-Fc versus hLIFR-Fc, two-tailed unpaired Student’s t test. f Both hLIFR-Fc and eLIFR-Fc (bivalent and one-arm) compete LIF away from WT gp130, but the D25 mAb does not and appears to increase binding, perhaps due to complex stabilization or more avid LIF binding. P = 0.02, ** P = 0.006, or *** P < = 0.0002 versus hLIFR-Fc or eLIFR-Fc (as indicated), two-tailed unpaired Student’s t test. For e and f , data are mean ± SD ( n ≥ 3 independent experiments).
Article Snippet: Recombinant hLIF-His (14890-H08H-20, Sino Biological Inc.), hLIF (untagged) (14890-HNAH-50, Sino Biological Inc.), mLIF-His (ABIN2215872, Antibodies-Online), hOSM-His (10425-H08H-20, Sino Biological Inc.), and hCTF-1-Fc (16013-H01H-20, Sino Biological Inc.) were purchased for use.
Techniques: Incubation, Two Tailed Test, Recombinant, Binding Assay