人Fcα/μR和Poly Immunoglobulin Receptor多克隆抗体的制备及鉴定

Preparation and purification of polyclonal antibodies against human Fcα/μR and the polymeric immunoglobulin receptor

目的:制备抗人Fcα/μR和Poly Immunoglobulin Receptor(pIgR)的多克隆抗体。方法:在Escherichia coli中分别表达人Fcα/μR的EC12和pIgR的D1结构域,均以包含体形式存在。从SDS-PAGE中切胶纯化,免疫新西兰兔,制备多克隆抗体。ELISA法检测抗体效价,Western blot法检测抗体的特异性,抗原柱吸收交叉和纯化多抗。结果:抗体滴度分别是1∶6 4000和1∶25 6000,经Western blot检测特异性良好,抗人Fcα/μR和pIgR的多抗相互没有交叉。结论:在E.coli中成功表达了人Fcα/μR的EC12结构域和pIgR的D1结构域,并制备了无交叉的多克隆抗体,为进一步研究其功能和相互关系奠定基础。

Objective：To generate rabbit anti-human Fcα/μR and Poly Immunoglobulin Receptor（pIgR） polyclonal antibodies.Methods：The EC12 domain of human Fcα/μR and the D1 domain of pIgR were expressed as including bodies in Escherichia coli.The purified proteins were used to immunize rabbits to generate polyclonal antibodies.The immune sera were characterized by ELISA and Western blot analysis,then absorbed with cross-action and purified by antigen linked beads.Results：The titers of antiserum were 1∶6 4000 and 1∶25 6000,separately.There was little cross-reaction between the two immune sera.Conclusion：The polyclonal antibodies against human Fcα/μR and pIgR without cross-reaction were successfully generated and could be useful tools to study the relationship between the two receptors.