Surugiu I, Danielsson B, Ye L, Mosbach K, Haupt K
Lund University, Department of Pure and Applied Biochemistry, Chemical Center, Sweden.
Anal Chem. 2001 Feb 1;73(3):487-91. doi: 10.1021/ac0011540.
An imaging assay analogous to competitive enzyme immunoassays has been developed using a molecularly imprinted polymer instead of an antibody. The antigen 2,4-dichlorophenoxyacetic acid (2,4-D) was labeled with tobacco peroxidase, and the chemiluminescence reaction of luminol was used for detection. Microtiter plates (96 or 384 wells) were coated with polymer microspheres imprinted with 2,4-D, which were fixed in place by using poly(vinyl alcohol) as glue. In a competitive mode, the analyte-peroxidase conjugate was incubated with the free analyte in the microtiter plate, after which the bound fraction of the conjugate was quantified. After addition of the chemiluminescent substrates, light emission was measured in a high-throughput imaging format with a CCD camera. Calibration curves corresponding to analyte concentrations ranging from 0.01 to 100 microg/mL were obtained.
已开发出一种类似于竞争性酶免疫测定的成像分析方法，该方法使用分子印迹聚合物代替抗体。抗原2,4-二氯苯氧乙酸（2,4-D）用烟草过氧化物酶标记，并利用鲁米诺的化学发光反应进行检测。用印有2,4-D的聚合物微球包被微量滴定板（96孔或384孔），使用聚乙烯醇作为胶水将其固定到位。在竞争模式下，将分析物-过氧化物酶偶联物与微量滴定板中的游离分析物一起孵育，然后对偶联物的结合部分进行定量。加入化学发光底物后，用CCD相机以高通量成像形式测量发光。获得了对应于浓度范围为0.01至100μg/mL的分析物的校准曲线。
