Xiao Qian, Zheng Yizhi, Liu Jing, Wang Suiping, Feng Bo
College of Chemical Engineering, Xiangtan University, Xiangtan 411105, Hunan Province, China.
College of Chemical Engineering, Xiangtan University, Xiangtan 411105, Hunan Province, China.
Biologicals. 2017 May;47:46-51. doi: 10.1016/j.biologicals.2017.02.008. Epub 2017 Mar 8.
In this study, we designed a comprehensive strategy for the ultrasensitive detection of alpha fetoprotein (AFP) with high specificity using gold nanoparticle (AuNP)-based enzyme-linked immunosorbent assay (ELISA). A dual-film modified probe was synthesized by coating AuNPs with horseradish peroxidase (HRP) on its surface. Anti-AFP monoclonal antibody (McAb) was immobilized on the surface of the enzyme using glutaraldehyde cross-linking method. AuNPs, employed as support for the immobilization of HRP. HRP was used not only as the enzymatic-amplified tracer but also as a bridge for loading McAb. The limit of detection was 2 ng mL. The developed probes can provide an alternative approach with high sensitivity and a simple process similar to that of the traditional HRP-McAb based ELISA for the ultrasensitive detection of AFP in serum.
在本研究中，我们设计了一种综合策略，利用基于金纳米颗粒（AuNP）的酶联免疫吸附测定（ELISA）对甲胎蛋白（AFP）进行高特异性的超灵敏检测。通过在其表面包被辣根过氧化物酶（HRP）的AuNP合成了一种双膜修饰探针。使用戊二醛交联法将抗AFP单克隆抗体（McAb）固定在酶的表面。AuNP用作固定HRP的载体。HRP不仅用作酶促放大示踪剂，还用作负载McAb的桥梁。检测限为2 ng/mL。所开发的探针可为血清中AFP的超灵敏检测提供一种具有高灵敏度且过程简单的替代方法，类似于基于传统HRP-McAb的ELISA。
