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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：1324 下载量：349
| 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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