Anti-human il-33 neutralizing autoantibody and preparation method and application thereof
A technology for autoantibodies and autoimmune diseases, applied in the field of anti-human IL-33 neutralizing autoantibodies and its preparation, can solve the problems of unknown importance and unsatisfactory
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Embodiment 1
[0053] Example 1 Preparation of anti-human IL-33 neutralizing autoantibodies by genetic engineering
[0054]Three main IL-33 epitope peptides were designed and synthesized by antigen epitope prediction software (DNAStar Protean software, SOPMA and the Bepi Pred2.0). The results are shown in Table 1. Through enzyme-linked immunosorbent assay (ELISA), it was found that serum samples from patients with positive anti-IL-33 antibodies could recognize the three IL-33 epitope peptides, which proved that the three IL-33 epitope peptides had high antigenicity and immunogenicity, can induce IL-33 antibody production. Therefore, these three IL-33 epitope peptides can be used as antigens to detect the level of anti-human IL-33 neutralizing autoantibodies in the blood of patients, and can also be used to activate and expand IL-33-specific B cells to produce anti-human IL-33. Human IL-33 neutralizing autoantibodies can also be used to obtain IL-33-specific B cells from blood by flow cytome...
Embodiment 2
[0059] Example 2 Induction of anti-human IL-33 neutralizing autoantibodies in vitro
[0060] Human peripheral blood mononuclear cells (PBMC) can be induced to produce specific antibodies by in vitro antigen immunization, which is a method for producing human-specific antibodies [7]. We use and improve this in vitro immunization method to produce anti-human IL-33 medium. and sex autoantibodies. Briefly, PBMCs were isolated from healthy donors negative for anti-human IL-33 neutralizing autoantibodies and immunosuppressive cells were depleted with Leu-leu methyl ester hydrobromide (LLME). Then three IL-33 epitope peptides (Table 1) were mixed with TLR9 ligand CpG-ODN (1 μM), cytokine IL-2 (50ng / ml), IL-4 (50ng / ml), respectively, in vitro After 7 days of immune cells, the supernatant was collected to obtain anti-human IL-33 neutralizing autoantibodies.
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