基于铜聚合物的荧光免疫分析用于 -苯二胺的催化氧化反应

Fluorescent Immunoassay with a Copper Polymer as the Signal Label for Catalytic Oxidation of -Phenylenediamine.

机构信息

College of Chemistry and Chemical Engineering, Pingdingshan University, Pingdingshan 467000, China.

College of Chemistry and Chemical Engineering, Henan University of Technology, Zhengzhou 450001, China.

出版信息

Molecules. 2022 Jun 8;27(12):3675. doi: 10.3390/molecules27123675.

This work suggested that Cu ion coordinated by the peptide with a histidine (His or H) residue in the first position from the free N-terminal reveals oxidase-mimicking activity. A biotinylated polymer was prepared by modifying His residues on the side chain amino groups of lysine residues (denoted as K) to chelate multiple Cu ions. The resulting biotin-poly-(K-Cu) polymer with multiple catalytic sites was employed as the signal label for immunoassay. Prostate specific antigen (PSA) was determined as the model target. The captured biotin-poly-(K-Cu) polymer could catalyze the oxidation of -phenylenediamine (OPD) to produce fluorescent 2,3-diaminophenazine (OPDox). The signal was proportional to PSA concentration from 0.01 to 2 ng/mL, and the detection limit was found to be eight pg/mL. The high sensitivity of the method enabled the assays of PSA in real serum samples. The work should be valuable for the design of novel biosensors for clinical diagnosis.

这项工作表明，在自由 N 端第一个位置由组氨酸（His 或 H）残基配位的 Cu 离子显示出氧化酶模拟活性。通过修饰赖氨酸（K）侧链氨基上的组氨酸残基来螯合多个 Cu 离子，制备了一种生物素化聚合物（表示为 K-Cu）。具有多个催化位点的生物素-聚（K-Cu）聚合物被用作免疫测定的信号标记物。前列腺特异性抗原（PSA）被确定为模型靶标。捕获的生物素-聚（K-Cu）聚合物可以催化 -苯二胺（OPD）的氧化生成荧光 2,3-二氨基吩嗪（OPDox）。信号与 0.01 至 2ng/mL 范围内的 PSA 浓度成正比，检测限发现为 8pg/mL。该方法的高灵敏度使得能够在实际血清样品中测定 PSA。这项工作对于设计用于临床诊断的新型生物传感器应该是有价值的。