| 李爱美,练玉银,王家骥.重组早孕因子与天然早孕因子的免疫交叉反应[J].,2018,(4):616-620 |
| 重组早孕因子与天然早孕因子的免疫交叉反应 |
| The Cross Reaction of the Recombinant Early Pregnancy Factor and the Native Early Pregnancy Factor |
| 投稿时间：2017-05-13 修订日期：2017-06-08 |
| DOI：10.13241/j.cnki.pmb.2018.04.004 |
| 中文关键词: 早孕因子 重组 天然 免疫交叉反应 |
| 英文关键词: EPF Recombination Native Cross reaction |
| 基金项目:广东省中医药局科研项目（20162101）；广州市教育科学规划课题（1201533519） |
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| 中文摘要: |
| 摘要 目的：探讨细胞表达的重组早孕因子(CHO-EPF)、大肠杆菌表达的重组早孕因子(BL21-EPF)和天然早孕因子(Native Early Pregnancy Factor，nEPF)之间的免疫交叉反应，寻找制备EPF抗体比较理想的免疫原。方法：采用CHO-EPF、nEPF作为免疫原分别免疫BALB/c小鼠，制备CHO-EPF多抗和nEPF多抗；结合本实验室储备的BL21-EPF鼠单克隆抗体，运用SDS-PAGE、Western blotting及ELISA等方法对nEPF、CHO-EPF、BL21-EPF之间的免疫交叉反应进行检测。结果：三种来源的EPF诱导抗体的效价比较无显著性差异。原核表达BL21-EPF与nEPF诱导的抗体中BL21-EPF单抗能识别nEPF中26 ku和52 ku组分，且抗nEPF多抗也能与BL21-EPF中10 ku组分反应；真核表达CHO-EPF与nEPF诱导的抗体中CHO-EPF多抗能识别nEPF中10 ku和26 ku组分，而nEPF抗体不能与CHO-EPF反应；原核表达BL21-EPF与真核表达CHO-EPF诱导的抗体中CHO-EPF多抗能识别BL21-EPF中10 ku片段，而BL21-EPF单抗不能与CHO-EPF反应。结论：真核源性CHO-EPF、原核源性BL21-EPF、人源性nEPF之间存在一定的交叉反应，在抗体制备过程中用rEPF替代nEPF作为免疫原是可行的。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the effect of EPF from different sources on antibody titers and the cross reaction between antibodies，and to clarify whether rEPF can replace human EPF as immunogen in the preparation of EPF antibody. Methods: Using the antigens which prepared with three kinds of different methods to immunize BALB/c mice. 3 kinds of antibodies，anti-CHO-EPF, BL21-EPF and nEPF were produced, determine their titer and the optimal antigen antibody concentration. The immunogenicity of EPF from different sources was evaluated by SDS-PAGE electrophoresis, Western blotting and ELISA. Results: Interaction experiments showed that the antibodies prepared from three kinds of EPF can identify native EPF.CHO-EPF antibodies recognize BL21-EPF and native EPF， but neither BL21-EPF nor native EPF antibodies do not recognize CHO-EPF. No significant difference in the titer of EPF antibody between the three sources. Conclusion: This study shows that eukaryotic EPF， prokaryotic EPF and human EPF have similar structures and slight differences. The antibody titers of EPFs vary from one source to another and there is a certain cross reaction between antibodies. And it is possible to demonstrate that it is feasible to replace nEPF as an immunogen with rEPF during EPF antibody preparation. |
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