【摘要】 以玻碳电极为工作电极研究了邻联茴香胺 ( ODA)为底物微分脉冲伏安法测定辣根过氧化物酶 ( HRP)及其标记物的方法。 HRP能够催化 H2 O2 氧化 ODA,其反应产物在玻碳电极上 - 0 .31 V ( vs.Ag/Ag Cl)左右被还原产生一个灵敏的还原峰 ,还原峰电流随着酶浓度的增大而增大 ,借助此还原电流可以测定 HRP,并可用于以 HRP为标记物的酶免疫分析。对酶催化反应条件和酶催化反应产物的测定条件进行了详细的研究 ,在最佳实验条件下测定游离 HRP的线性范围是 4.0× 1 0 - 10 ～ 6.0× 1 0 - 8g· m L- 1,检测限为3.3× 1 0 - 10 g· m L- 1;测定游离的酶标记物 ( Ig G- HRP) ,稀释范围为 1∶ 2 0 0 0～1∶ 1 0 0 0 0 0 0 ,最大稀释比为 1∶ 1 0 0 0 0 0 0。更多还原

【Abstract】 A voltammetric detection of horseradish peroxidase(HRP)and enzyme labeled IgG using glass carbon electrode (GCE) was investigated based on o-Dianisidin (ODA)-H 2O 2-HRP coupling reaction system. The product of ODA oxidized by H 2O 2 with presence of HRP can be reduced at -0.31 V (vs.Ag/AgCl) on glass carbon electrode and the differential pulse voltammetry (DPV) was chosen to detect the enzymatic-generated product. The conditions for HRP enzymatic reaction and electrochemical detection were carefully studied with a CHI 832 electrochemical analyzer. The reductive peak current is linear with the HRP concentration from 4.0×10 -10 to 6.0×10 -8g·mL -1 and the detection limit for HRP is 3.3×10 -10 g·mL -1.The highest dilution ratio of free IgG-HRP is 1∶1000000 This system can be further used in HRP-labeled electrochemical enzyme-linked immunoassay.更多还原