军事医学2017，Vol.41Issue(3)：184-189,6.DOI:10.7644/j.issn.1674-9960.2017.03.006
抗人Her2抗原羊驼纳米抗体的筛选与鉴定
Generation of an alpaca derived nanobody recognizing human Her2 antigen
王兰东 1刘枫 2成岩 2李大磊 3韩冰 3张淑敏 1冯东晓3
作者信息
- 1. 滨州医学院药学院,山东烟台 264000
- 2. 山东绿叶制药有限公司,山东烟台 264000
- 3. 山东国际生物科技园发展有限公司,山东烟台 264000
- 折叠
摘要
Abstract
Objective To obtain alpaca single domain antibody targeting Her2.Methods An alpaca was immunized with human recombination Her2 protein mixed with Freund's adjuvant.Total RNA was extracted from the alpaca's blood and was used to synthesize first strand cDNA.Single domain antibody variable region (VHH) gene of the alpaca was amplified by PCR and cloned into pMES4 vector for library construction.After screening, E.coli BL21 (DE3) was transformed with selected clones and was induced with IPTG for the expression of recombinant proteins.The nanobody was purified by nickel ion affinity chromatography column.The affinity of the nanobodies to Her2 was tested.Results After the second round of screening, two antibody clones were selected, H3 and H5.The affinity of H5 was 8.106×10-10mol/L.Histochemistry results showed that H5 could recognize Her2 antigen in breast tumor tissue.Conclusion An Her2 specific nanobody derived from alpaca is obtained through phage display library screening, which can recognize human Her2 antibody in human breast tumor tissue.关键词
Her2/纳米抗体/噬菌体展示技术/乳腺肿瘤/诊断Key words
Her2/nanobody/phage display technology/breast neoplasms/diagnosis分类
医药卫生引用本文复制引用
王兰东,刘枫,成岩,李大磊,韩冰,张淑敏,冯东晓..抗人Her2抗原羊驼纳米抗体的筛选与鉴定[J].军事医学,2017,41(3):184-189,6.
