中国免疫学杂志2024，Vol.40Issue(7)：1501-1506,6.DOI:10.3969/j.issn.1000-484X.2024.07.026
IGFBP-3抗原的真核表达分析及校准品制备评估
Eukaryotic expression analysis and calibration materials evaluation of IGFBP-3 antigen
摘要
Abstract
Objective:To observe the expression differences of IGFBP-3 antigen in different eukaryotic cells,and the optimal antigen was screened for calibration materials evaluation to develop IGFBP-3 antigen diagnostic kit.Methods:Human IGFBP-3 gene was respectively cloned into Pichia pastoris expression vector pPIC9K and mammalian expression vector pCMV,and then transformed into Pichia pastoris GS115,HEK293 cell and CHO cell for gene expression.Activity analysis and purification of the expressed prod-ucts were performed with Ni-NTA and SDS-PAGE,and calibrating performance of the purified antigen was analyzed with IGFBP-3 test kit.Results:IGFBP-3 antigen was easily degradable during the expression process,and only the HEK293 cells enabled to obtain the full-length antigen,and reactivity and stability of the calibration products using antigen expressed in HEK293 cells could meet the calibration material requirements for diagnostic kits,laying a solid foundation for further development of IGFBP-3 diagnostic kits.Conclusion:IGFBP-3 expressed by HEK293 cells is more suitable for calibration materials preparation.关键词
IGFBP-3抗原/真核表达/校准品Key words
IGFBP-3 antigen/Eukaryotic expression/Calibration materials分类
医药卫生引用本文复制引用
王岚,贾国超..IGFBP-3抗原的真核表达分析及校准品制备评估[J].中国免疫学杂志,2024,40(7):1501-1506,6.基金项目
2019年度河南省青年骨干教师项目(2019GGJS272). （2019GGJS272）
