Surugiu L, Svitel J, Ye L, Haupt K, Danielsson B
Department of Pure and Applied Biochemistry, Chemical Center, Lund University, Sweden.
Anal Chem. 2001 Sep 1;73(17):4388-92. doi: 10.1021/ac0101757.
A flow injection competitive assay analogous to enzyme immunoassays has been developed using a molecularly imprinted polymer instead of the antibody. A glass capillary was modified by covalently attaching an imprinted polymer to the inner capillary wall. The herbicide 2,4-dichlorophenoxyacetic acid was used as a model analyte. The analyte was labeled with tobacco peroxidase, and chemiluminescence was used for detection in combination with a photomultiplier tube or a CCD camera. In a competitive mode, the analyte-peroxidase conjugate was passed together with the free analyte through the polymer-coated capillary mounted in a flow system. After a washing step, the chemiluminescent substrate was injected and the bound fraction of the conjugate was quantified by measuring the intensity of the emitted light. Calibration curves corresponding to analyte concentrations ranging from 0.5 ng mL(-1) to 50 microg mL(-1) (2.25 nM-225 microM) were obtained. A lowered detection limit by 2 orders of magnitude was obtained when detection was done in discontinuous mode and the chemiluminescence light was conducted inside the photomultiplier tube by an optical fiber bundle, thus yielding a dynamic range of 5 pg mL(-1)-100 ng mL(-1) (22.5 pM-450 nM).
已开发出一种类似于酶免疫测定的流动注射竞争测定法，该方法使用分子印迹聚合物替代抗体。通过将印迹聚合物共价连接到玻璃毛细管内壁对其进行修饰。以除草剂2,4-二氯苯氧乙酸作为模型分析物。分析物用烟草过氧化物酶标记，并结合光电倍增管或电荷耦合器件相机使用化学发光进行检测。在竞争模式下，分析物 - 过氧化物酶偶联物与游离分析物一起通过安装在流动系统中的聚合物涂层毛细管。经过洗涤步骤后，注入化学发光底物，并通过测量发射光的强度对偶联物的结合部分进行定量。获得了对应于浓度范围为0.5 ng mL(-1)至50 μg mL(-1)（2.25 nM - 225 μM）的分析物的校准曲线。当以不连续模式进行检测且化学发光光通过光纤束在光电倍增管内部传导时，检测限降低了2个数量级，从而得到5 pg mL(-1)-100 ng mL(-1)（22.5 pM - 450 nM）的动态范围。
