[Preparation and identification of mouse monoclonal antibodies against human IgE]

Objective To prepare the high-affinity and high-specificity mouse anti-human IgE monoclonal antibody (mAb) as a candidate immunosorbent. Methods BALB/c mice were immunized using recombinant antigen IgECepsilon2-4. Coated ELISA plate with IgECepsilon2-4 was used to screen positive cell lines by indirect ELISA, then coated ELISA plate with natural IgE, IgG, IgM, IgA, IgD to detect the affinity and specificity of serum-free culture supernatant of positive hybridoma cells with natural IgE. The cell line stably secreting specific anti-IgE monoclonal antibodies was expanded and inoculated into the abdominal cavity of BALB/c mice to prepare mAbs. The secreted mAbs were identified by ELISA kit followed by the identification of mAb subtypes. Results The 29 positive hybridoma cells were obtained after five cell fusions, of which 11 strains had strong affinity with natural IgE and 2 strains did not cross-react with other immunoglobulins 3E9 and 7B4. Conclusion The study successfully prepared mAbs against human IgE with high titer, affinity and specificity.