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环境水样中Cr6+酶联免疫检测方法的建立

发表时间:2012-09-15  浏览量:918  下载量:284
全部作者: 邹军辉,唐勇,钟辉,宋建勋,欧子强
作者单位: 暨南大学广东省分子免疫和抗体工程重点实验室
摘 要: 以抗Cr3+-EDTA单克隆抗体为基础,建立优化的间接竞争酶联免疫检测(enzyme-linked immunosorbent assay,ELISA)方法和样品前处理方法,用于环境水样中Cr6+的检测。分别配制不同pH值及不同离子强度的PBS保温液,优化筛选ELSIA的最佳反应pH值与离子强度。前处理过程中,通过调节样品pH值,沉淀去除水样原有Cr3+;优化各项还原条件使Cr6+还原为Cr3+,调节pH值进行ELISA检测,通过检测出的Cr3+含量计算原有Cr6+的含量。结果表明:当ELISA保温液pH值为7.4、离子强度为0.15 mol/L时,ELISA的灵敏度(IC50)最佳,为5.9 μg/L,检测限为0.35 μg/L,线性范围为0.35~50 μg/L. 当还原剂为60 mg/L NaHSO3溶液、还原pH值为3.0、反应时间为15 min时样品前处理效率最高。以结合最优样品前处理方法的ELISA检测实际水样并与等离子体发射光谱法(inductively coupled plasma, ICP)检测结果比较,得到2种方法检测结果数据的相关系数达0.988 7,表明研究建立的ELISA方法检测结果可靠。加标实验结果显示该ELSIA方法变异系数小于12%,说明该方法具有良好的重复性和精密度,可用于环境水样Cr6+的监测。
关 键 词: 免疫生物化学;Cr6+;酶联免疫检测;单克隆抗体;环境水样
Title: Development of enzyme-linked immunoassay for Cr6+ detection in environmental water samples
Author: ZOU Junhui, TANG Yong, ZHONG Hui, SONG Jianxun, OU Ziqiang
Organization: Guangdong Province Key Laboratory of Molecular Immunology and Antibody Engineering, Jinan University
Abstract: An optimal indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed using a new monoclonal antibody against Cr3+-EDTA. The pretreatment procedure was also established. Reaction conditions, such as pH and ionic strength, were studied to optimize the immunoassay. In pretreatment procedure, the Cr3+ existent in samples was precipitated by adjusting pH and the reducing conditions of Cr6+ were researched completely, the concentration of Cr6+ in samples were calculated as Cr3+ detected by ELISA. Results showed that the ELISA had the most sensitive to soluble Cr3+-EDTA when pH was 7.4 and ionic strength was 0.15 mol/L, the IC50 and limit of detection was 5.9 μg/L, 0.35 μg/L, respectively, and the liner range was 0.35-50 μg/L. The collected supernatant containing Cr6+ was mixed with 60 mg/L of NaHSO3 solution, the Cr6+ was reduced to Cr3+ at pH 3.0 and reacted for 15 min. Finally, ELISA was used to detect the concentration of Cr6+ in samples. The correlation coefficient between the ELISA and ICP was 0.988 7 and the coefficient of variation was below 12%, indicating that the immunoassay was reliable and precise, and it was suitable to detect water samples.
Key words: immunobiochemistry; Cr6+; enzyme-linked immunosorbent assay; monoclonal antibody; environmental water samples
发表期数: 2012年9月第17期
引用格式: 邹军辉,唐勇,钟辉,等. 环境水样中Cr6+酶联免疫检测方法的建立[J]. 中国科技论文在线精品论文,2012,5(17):1643-1648.
 
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