中成药2024，Vol.46Issue(4)：1214-1219,6.DOI:10.3969/j.issn.1001-1528.2024.04.023
基于特征多肽抗原的阿胶基原鉴定
Origin identification of Asini Corii Colla based on characteristic polypeptide antigens
摘要
Abstract
AIM To identify the origin of the raw materials of Asini Corii Colla by preparing polyclonal antibodies with characteristic polypeptides as antigens.METHODS The characteristic polypeptides were screened by literature research and database comparison.Balb/c mice were immunized with KLH-conjugated polypeptides to prepare polyclonal antibodies.Then,the production,titers and specificities of the polyclonal antibodies were analyzed and verified by ELISA,dot blot and Western blot.RESULTS All the five characteristic polypeptides had T cell and B cell epitopes,with hydrophilicity of(-1-0),and most of the secondary structures were random curling.After the second booster immunization,the titers of Ab-ECS1,Ab-ECS3,Ab-ECS4 and Ab-ECS5 were 1 ∶ 6 400,1 ∶ 400,1 ∶ 12 800 and 1 ∶ 800,respectively.In addition,there was no cross-reaction between the prepared polyclonal antibodies,and the binding of the polyclonal antibody Ab-ECS3 to the donkey skin and its counterfeit proteins showed a large difference.CONCLUSION The polyclonal antibodies prepared with the characteristic polypeptides as hapten can be used to identify the authenticity of donkey skin by Western blot,which provides a reference for the identification of the origin of gelatinous Chinese medicine.关键词
阿胶/特征多肽/多肽抗原/多克隆抗体/Western blot/基原鉴定Key words
Asini Corii Colla/characteristic polypeptides/polypeptide antigens/polyclonal antibodies/Western blot/origin identification分类
医药卫生引用本文复制引用
袁陈婷,杨容,王立玮,景凌洁,陈璐阳,加羊卓玛,孙小祥,孙锦秀,杨欢..基于特征多肽抗原的阿胶基原鉴定[J].中成药,2024,46(4):1214-1219,6.基金项目
国家自然科学基金项目(81773855) （81773855）
北京康盟慈善基金会卫生健康发展促进项目(KM-ZLGJ-009) （KM-ZLGJ-009）
南通市基础科学研究和民生科技计划项目(JCZ2022031) （JCZ2022031）
镇江市科技计划项目(SH2021057) （SH2021057）
