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. 2017 Dec 26;9(4):5197–5207. doi: 10.18632/oncotarget.23708

Figure 1. Specificity, internalization and affinity of promiximab.

Figure 1

(A) Western blot detected CD56 expression in small cell lung cancer cell lines cell lysates including CD56 extracellular domain (CD56 ECD), NCI-H526, NCI-H524, NCI-H69, NCI-H128, NCI-H446 using promiximab antibody. (B) Western blot was required for detection of CD56 extracellular domain on the condition of deglycosylation or non-deglycosylation using promiximab antibody. (C) Identification of cross reactivity of chimeric antibody with human NK (hNK) cell lysates, human spleen (hSpleen) cell lysates, mouse spleen (mSpleen) cell lysates by western blot, CD56 ECD as control. (D) Binding capacity of promiximab to small cell lung cancer cell lines were detected by flow cytometric analyses and SCLC cell lines incubated with PBS (purple), Promiximab (green), respectively. (E) Internalization of promiximab in CD56-positive small cell lung cancer cell lines were also assessed by flow cytometric analyses, cell lines were incubation with PBS (purple, 4°C, 3 h), promiximab (green, 4°C, 3 h), promiximab (fuchsia, 37°C, 3 h). (F) Biolayer interferometry binding assay were used for monitoring the interaction kinetics between CD56 ECD and different concentrations of promiximab as described in Materials and Methods. Three independent experiments were conducted and similar results were obtained.