IL-18 binding proteins
A technology for binding proteins and binding domains, which can be used in antifungal agents, antibacterial agents, sugar derivatives, etc., and can solve problems such as inability to signal transduction
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Embodiment 1
[0285] Example 1: Preparation and characterization of recombinant IL-18
Embodiment 11
[0286] Example 1.1: IL-18 biological activity determination test
[0287] In Example 1 and Example 2, unless otherwise specified, the following test was used to determine the biological activity of IL-18.
Embodiment 11A
[0288] Example 1.1.A: KG-1 bioassay
[0289] KG-1 (ATCC#CCL-246) is a human bone marrow mononuclear cell line that constitutively expresses low-level functional IL-18 receptors. The use of TNF therapy up-regulates the IL-18R alpha and beta subunits of the functional IL-18 receptor on these cells. By ELISA, TNF-treated KG-1 cells were incubated with recombinant human IL-18 (rhu-IL-18) for KG-1 bioassay to determine the level of IL-18-induced human IFNγ production (Konishi, K ., et al. (1997) J. Immunol. Methods 209:187-191). The KG-1 bioassay was used to determine the neutralizing ability of IL-18 antagonists. For example, anti-IL-18 antibodies are incubated with different concentrations of rhu-IL-18, and then incubated with TNF-treated KG-1 cells in a 96-well plate at 37°C for 16-18 hours. The supernatant was collected and the human IFNγ level was determined by ELISA. The test measured IL-18 antagonists as low as 4×10 -11 To 6×10 -11 M's IC 50 value
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