紫外吸收光谱积分法分析蛋白质浓度-以碱性磷酸酶为例  

Quantitative Analysis of Protein Concentration by Absorption Peak Integration Method of UV Spectroscopy——Taking Alkaline Phosphatase as an Example

作　　者：朱元荣[1,2] 吴丰昌[1] 林樱[3] 

机构地区：[1]中国环境科学研究院环境基准与风险评估国家重点实验室,北京100012 [2]北京师范大学水科学研究院,北京100875 [3]环境保护部环境工程评估中心,北京100012

出　　处：《光谱学与光谱分析》2013年第7期1845-1849,共5页Spectroscopy and Spectral Analysis

基　　金：国家重点基础研究发展(973计划)项目(2008CB418200);国家自然科学基金项目(41261140337;40973090)资助

摘　　要：利用矿物(针铁矿,蒙脱石)和太湖沉积物吸附碱性磷酸酶(alkaline phosphatase,APase),测定吸附后上清液中剩余碱性磷酸酶浓度时发现其紫外吸收光谱发生了变化,利用传统280nm处紫外吸收法无法直接准确测定其浓度值。基于对碱性磷酸酶252～305nm处吸收峰面积积分方法可以消除影响,并准确分析测定碱性磷酸酶浓度。其测定结果与考马斯亮蓝法测定结果进行比较,表明了该方法可以方便,快速和准确地测定此类实验中碱性磷酸酶浓度。同时,该方法还可以扩展至其他蛋白质的定量分析,甚至其他类似实验中,一定程度上克服传统方法应用单波长进行定量分析中存在的易受干扰的缺点。Adsorption of alkaline phosphatase （APase） on minerals （goethite, montmorillonite） and sediments from Taihu Lake was studied. However, the concentration of alkaline phosphatase in the supernatant cannot be analyzed by the adsorption at 280 nm due to that the UV spectroscopy was changed. Quantitative analysis of alkaline phosphatase in the supernatant by the ab- sorption peak （252--305 nm） integration method of UV spectroscopy was developed. This method determined the concentrations of alkaline phosphatase well. Compared to the results of Bradford, the absorption peak integration method can determine the concentrations of supernatant fast, conveniently, and accurately. This method can also be applied to other protein solution analy- sis and similar experiments. The drawbacks of traditional single wavelength method （280nm） were overcome to some extent based on the method of absorption peak integration.

关 键 词：紫外吸收光谱 峰面积积分 蛋白质 碱性磷酸酶 测定 

分 类 号：O657.3[理学—分析化学]