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[克百威免疫亲和色谱法的研究]

[Studies of immunoaffinity chromatography for carbofuran].

作者信息

Liu Shuzhao, Wei Linhong, Xu Weina

机构信息

College of Environmental Science and Engineering, Yangzhou University, Yangzhou 225009, China.

出版信息

Se Pu. 2005 Mar;23(2):134-7.

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 microg/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.

摘要

将纯化的抗克百威抗体与羰基二咪唑(CDI)活化的琼脂糖凝胶CL-4B偶联,合成用于克百威特异性免疫亲和色谱(IAC)柱的免疫吸附剂。IAC的条件优化如下:pH 7.2的磷酸盐缓冲液(PB)用作平衡和吸附介质,甲醇-水(60:40,v/v)用作洗脱剂。结果表明,动态柱容量高达1.58 mg/L床体积。当标准样品溶液中克百威的初始浓度低于2 μg/L时,IAC的富集效率超过167倍。加标的河水经IAC净化和富集,洗脱液中的克百威采用酶联免疫吸附测定(ELISA)法测定。在加标水平为0.1 mg/L时,河水中克百威的平均回收率为89.8%,相对标准偏差为4.8%。同时,采用高效液相色谱(HPLC)对洗脱液进行测定,HPLC结果与ELISA结果相关性良好。成功建立了克百威的IAC方法。

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