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. 2017 Jan 17;9(3):506–520. doi: 10.1080/19420862.2016.1274844

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Figure 5. — Expression and purification of human and murine GpL(_CT-LC)-IgG variants of 18D1. (A, B) Purified GpL_(CT-LC)-18D1-IgG2, GpL_(CT-LC)-18D1-mIgG1 and GpL_(CT-LC)-18D1-mIgG2a (200 ng) were analyzed by SDS-PAGE (A) and gel filtration (B). (C) Equilibrium binding studies with the various novel GpL_(CT-LC)-18D1-IgG variants and HT1080 and HT1080-Fn14-KO cells were performed as in Fig. 2B to determine their affinity for cell expressed Fn14. Shown is one representative experiment for each fusion protein. (D) Determination of the K_D values of 18D1-IgG2, 18D1-mIgG1 and 18D1-mIgG2a by competition binding experiments with 100 ng/ml of GpL_(CT-LC)-18D1-IgG2, GpL_(CT-LC)-18D1-mIgG1 and GpL_(CT-LC)-18D1-mIgG2a.