基于生物膜干涉技术的单克隆抗体高通量定量检测方法的建立
DOI:
CSTR:
作者:
作者单位:

1.福建医科大学 免疫治疗研究院,福州350122;2.福建卫生职业技术学院 药学系,福州350101;3.福建医科大学 药学院,福州350122

作者简介:

通讯作者:

中图分类号:

R392.11; R394.2; R446.1

基金项目:

收稿日期: 2020|01|14基金项目: 福建省教育厅中青年教师教育科研项目(JT180169);福建省卫生计生委青年科研课题(2016|2|38)作者简介: 胡妍,女,实验师,医学硕士通讯作者: 刘全裕. Email: liuquanyu0597@163.com


Establishment of A High|throughput Quantitative Method for Monoclonal Antibody Based on Bio|layer Interference Technique
Author:
Affiliation:

1. Immunotherapy Institute, Fujian Medical University, Fuzhou 350122, China;2. Department of Pharmacy, Fujian Health College, Fuzhou 350101, China;3. Scholl of Pharmacy, Fujian Medical University, Fuzhou 350122, China

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    目的运用生物膜干涉(BLI)技术,建立高通量定量检测单克隆抗体浓度的方法。方法使用Octet Red 96分子相互作用分析仪,根据样品与protein A传感器结合后光相对干涉位移变化,检测抗体浓度,并对这种检测方法进行方法学考察。结果protein A传感器能特异性地与单克隆抗体结合,抗体工作标准品浓度在7.81~500 μg/mL与结合速率呈良好的回归关系,相关系数R2为0.999 9;检测限为0.6 μg/mL,定量限为2 μg/mL;回收率为98.90%~100.40%;重复性的相对标准偏差(RSD)为2.17%,中间精密度的RSD为2.52%;45 d内protein A传感器重复使用60次,阳性对照值的RSD为3.7%;再生液pH变化±0.2、平衡液吐温\|20比例变化±5%的RSD值为0.99%。检测温度与转盘速度的变化是影响样品定量检测的因素,检测结果与高效液相数据高度一致(R2=0.90)。结论运用BLI技术能实现高通量定量检测单克隆抗体浓度,该方法具有样品适应性强、分析速度快、准确度高的优势,可用于单克隆抗体细胞株的筛选、排序及蛋白表达的检测与监控

    Abstract:

    ObjectiveTo observe the difference in MRI image and ultra\|structure in cerebral artery occlusion (MCAO) model and the photochemical induced focal cerebral infarction model in the rat brain.MethodsThe model of MCAO and photochemical induced focal cerebral infarction rats were established separately, and the ischemic area and cortical vessels distribution were observed by small animal magnetic resonance imaging system, and the ultra\|structural changes of the two models were compared with transmission electron microscopy (TEM).ResultsThe infarct area of MCAO model rat was large and unstable, the cell structure was destroyed seriously, while the vascular deformation was remarkable with few neovascularization.Whereas the infarct area of brain by photochemical induced method was small and stable, the cell structure damage was not serious, and there were more new blood vessels.ConclusionThe two model methods have the advantages and disadvantages, which can provide a practical tool for the protection of blood vessel injury after cerebral infarction and the thrombolytic therapy, and they may be used to select the appropriate model according to the demand of scientific research

    参考文献
    相似文献
    引证文献
引用本文

胡妍, 罗安, 车耀健, 刘全裕.基于生物膜干涉技术的单克隆抗体高通量定量检测方法的建立[J].福建医科大学学报自然版,2020,(3):139-143

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期:
  • 出版日期:
文章二维码