色谱

色谱

• 研究报告 • 上一篇    下一篇

克百威的免疫亲和色谱分析研究

刘曙照,韦林洪,徐维娜   

  1. College of Environmental Science and Engineering, Yangzhou University, Yangzhou 225009, China
  • 收稿日期:2004-08-23 修回日期:1900-01-01 出版日期:2005-03-30 发布日期:1988-12-25
  • 通讯作者: 刘曙照
  • 基金资助:
    国家自然科学基金资助项目(No.20177019).

Studies on Immunoaffinity Chromatography for Carbofuran

LIU Shuzhao, WEI Linhong, XU Weina   

  1. College of Environmental Science and Engineering, Yangzhou University, Yangzhou 225009, China
  • Received:2004-08-23 Revised:1900-01-01 Online:2005-03-30 Published:1988-12-25

PDF

817

被引次数

106

摘要： Sepharose CL-4B经碳酰二咪唑(CDI)活化后与纯化的克百威抗体共价偶联,合成了免疫亲和色谱(IAC)固定相,并用其制备了对克百威具有特异性亲和力的IAC柱。对IAC条件进行了优化,选择0.02 mol/L pH 7.2磷酸盐缓冲液(PB)作为吸附与平衡介质,60%（体积分数）甲醇水溶液作为洗脱剂。结果表明:在优化条件下,IAC柱对克百威的动态柱容量达1.58 mg/L。当标样溶液中克百威质量浓度低于 2　μg/L时,经IAC柱富集的效率高于167倍。在河水中按0.1 mg/L水平添加克百威标准品,经IAC柱分离富集,洗脱液采用包被抗体直接竞争酶联免疫吸附分析(ELISA)法检测,5次重复测定的平均回收率为89.8%,相对标准偏差为4.8%。同时采用高效液相色谱(HPLC)法测定洗脱液,与ELISA法测定结果基本一致。

关键词: 高效液相色谱, 克百威 , 酶联免疫吸附分析, 免疫亲和色谱

Abstract: The purified anti-carbofuran antibody was conjugated to carbonyl diimidazole (CDI)-activated Sepharose CL-4B to synthesize the immunosorbent for the immunoaffinity chromatographic (IAC) column specific to carbofuran. The conditions of IAC were optimized as follows: pH 7.2 phosphate buffer (PB) was used as equilibrium and adsorbent medium, and methanol-water (60∶40, v/v) as eluent. The results showed that the dynamic column capacity was up to 1.58 mg/L bed volume. The efficiency of enrichment of IAC was more than 167 times when the initial concentration of carbofuran in a standard sample solution was lower than 2 μg/L. The spiked river water was cleaned up and enriched by IAC, and carbofuran in eluate was determined by enzyme linked immunosorbent assay (ELISA). The average recovery of carbofuran from river water was 89.8% with the relative standard deviation of 4.8% at the spiked level of 0.1 mg/L. Meanwhile the eluate was determined by high performance liquid chromatography (HPLC), the results from HPLC correlated well with those from ELISA. The IAC method of carbofuran was successfully established.

Key words: carbofuran , enzyme-linked immunosorbent assay, high performance liquid chromatography, immunoaffinity chromatography