以CpG为佐剂的人IL-24多克隆抗体的制备

目的：采用在原核表达系统表达的人IL-24蛋白及人IL-24真核重组质粒免疫新西兰兔，制备兔抗人IL-24多克隆抗体。方法：利用IPTG诱导hIL-24在大肠杆菌中表达，纯化hIL-24重组蛋白，纯化后的蛋白经SDS—PAGE分析，同时提取hIL-24真核重组质粒，用以免疫新西兰兔，并以CpG为佐剂制备多克隆抗体。Western-blot鉴定抗体的特异性，ELISA法测定抗体效价。结果：人IL-24经IPTG诱导后可在大肠杆菌中大量表达，表达量占细菌总蛋白的30％，纯化后的蛋白纯度高；原核表达的重组蛋白和真核重组质粒免疫新西兰兔，通过抗体效价测定，获得了抗血清效价达1：640的多克隆抗体。结论：原核表达的hIL-24蛋白和hIL-24真核重组质粒均能刺激家兔产生抗体．其多克隆抗体的效价较高。

CpG-ODN is an adjuvant for preparation of the hIL-24 polyclonal antibody

Objective:To immunize rabbits with hIL-24 protein expressed in E.coli and eukaryotic recombinant plasmid to prepare hIL-24 polyclonal antibodies.Methods:The recombinant protein was achieved in E.coli through IPTG induction and was purified. The protein was analyzed by SDS-PAGE. Rabbits were immunized with hIL-24 protein and eukaryotic recombinant plasmid to prepare hIL-24 polyclonal antibodies.CpG-ODN was used as an adjuvant for preparation of the hIL-24 antibody.The antibodies were detected by western-blot and ELISA.Results:Human IL-24 could be expressed in E.coli after IPTG induction. The recombinant protein was about 30% of total expressed products in E.coli. Purified protein was achieved. The polyclonal antibodies were successfully prepared and had higher titter (1:640). Conclusion:HIL-24 recombinant protein expressed in E.coli and eukaryotic recombinant plasmid can stimulate antibody in rabbits.