Preparation and identification of monoclonal antibodies against human IgE

VernacularTitle:阻断型抗人IgE单克隆抗体的制备及其表位分析
Author: Jiaqi CHEN ; Aidong QU ; Jingye ZHU ; Haiwu HUANG
Publication Type:Journal Article
Keywords: Human IgE; Monoclonal antibody; Antigenic epitope
From:Chinese Journal of Microbiology and Immunology 2012;(10):845-849
CountryChina
Language:Chinese
Abstract: Objective To obtain monoclonal antibodies(McAb) against human IgE by recombinant IgE antigen.Methods BLAB/c mice were immunized with recombinant IgE.The spleen cells of immunized mice were used to prepare the McAb by hybridoma techniques.IgE-McAb was selected by indirect ELISA with culture supernatant of hybridoma cells.ELISA blocking assay were employed to identify the function of McAb.Alanine scanning mutagenesis were used to confirm the antigenic epitopes of McAb.Results A monoclonal antibody against IgE(178) was produced.The antigenic epitope of 178 was found to be spatially close to the receptor-binding site.Functional assay revealed that 178 could markedly inhibit IgE binding to receptors.Conclusion A hybridoma cell line secreting IgE-McAb stably was obtained by recombinant IgE antigen,and the IgE-McAb might block IgE binding to receptors via steric hindrance.