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Fig. 2 | Arthritis Research & Therapy

Fig. 2

From: The preclinical pharmacology of the high affinity anti-IL-6R Nanobody® ALX-0061 supports its clinical development in rheumatoid arthritis

Fig. 2

Inhibition profiles of ALX-0061 in an sIL-6R-based (a) and an mIL-6R-based (b) assay. ALX-0061 was pre-incubated with recombinant hIL-6 and recombinant hIL-6R, in the presence or absence of HSA, followed by the capture of hIL-6R on ELISA plates coated with a non-neutralizing anti-IL-6R mAb. IL-6 that remained in complex with IL-6R was detected using a biotinylated anti-IL-6 tool, and subsequent visualized with streptavidin-HRP. An example experiment for a neutralization experiment is presented in (a). Human TF-1 cells were pre-incubated with a dilution series of ALX-0061 in the presence of HSA, after which proliferation was induced with 2 ng/mL of IL-6. After 72 hours of incubation, cell proliferation was assessed by incorporation of 3H-thymidine. ALX-0061 did not induce proliferation in the absence of IL-6 (b). Symbols depict mean responses; error bars represent ± SD of triplicate samples within the experiment. ELISA: enzyme-linked immunesorbent assay; h: human; HRP: horseradish peroxidase; HSA: human serum albumin; IL-6: interleukin-6; IL-6R: IL-6 receptor; mAb: monoclonal antibody; mIL-6R: membrane IL-6R; OD: optical density; SD: standard deviation; sIL-6R: soluble IL-6R

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