抗EphA4多肽单克隆抗体的制备及初步应用

Preparation and analysis of monoclonal antibody against EphA4 peptide

目的:制备抗EphA4羧基端多肽的单克隆抗体,研究其免疫学特性。方法:体外合成EphA4羧基端多肽免疫小鼠,运用杂交瘤技术建立抗EphA4多肽单克隆抗体细胞株。通过免疫化学方法分析单抗的免疫学特性。结果:获得1株抗EphA4单克隆细胞,所分泌的抗体具有EphA4特异性,能识别人类、鸡与小鼠EphA4;可用于EphA4的ELISA、免疫沉淀、免疫印迹与免疫组织化学染色。结论:抗EphA4多肽单抗具备优良的免疫学特性,适用于多种免疫化学技术,是EphA4及其配体研究的重要工具。

Objective To prepare monoclonal antibody （MAb） against carboxy-terminus of EphA4 and to analyze the immunological characteristics and significance of the antibody. Methods Mice were immunized with a chemically synthesized peptide that had been conjugated to KLH （keyhole limpet hemocyamin）. A panel of antibodies specifically against EphA4 were obtained by hybridoma technique. The immunological properties and significance of the MAb were analyzed by immunological and immunochemistry techniques. Results A hybridoma cell line secreting anti-EphA4 MAb was established. The MAb reacted specifically with human, chicken and mouse EphA4, but did not cross-react with EphA5 and EphA7. The antibody could be used for immunochemical techniques such as ELISA, immunopreeipitation, Western blot and immunohistocnemistry. Conclusion The anti-EphA4 MAb generated by immunizing a synthetic peptide possesses excellent immunological properties. MAb can be applied for immunological and immunohistochemical purpose and will become an important tool in the study of EphA4 and its ligand.