Department of Pharmaceutical Engineering, School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou, 510006, People's Republic of China.
Faculty of Chemical Engineering and Light Industry, Guangdong University of Technology, Guangzhou, 510006, People's Republic of China.
Anal Bioanal Chem. 2022 Sep;414(23):6939-6946. doi: 10.1007/s00216-022-04261-7. Epub 2022 Aug 10.
Ferritin, widely present in liver and spleen tissue, is considered as a serological biomarker for liver diseases and cancers. The detection of ferritin may be an important tool in health diagnosis. In this study, 14 non-immunized chicken spleens were utilized to construct a single-chain fragment (scFv) phage library. After 4 rounds of panning, 7 unique clones were obtained. The optimal clone was further screened and combined with NanoLuc luciferase (Nluc) as a dual functional immunoprobe to bioluminescent enzyme immunoassay (BLEIA), which was twice as sensitive as its parental scFv-based double-sandwich enzyme-linked immunoassay (ds-ELISA). The cross-reactivity analysis revealed that the proposed methods were highly selective and suitable for clinical detection. To further verify the performance of the immunoassays, serum samples were tested by the proposed methods and a commercial ELISA kit, and there was a good correlation between the results. These results suggested that scFv fused with Nluc might be a powerful dual functional tool for rapid, practically reliable, and highly sensitive ferritin detection.
铁蛋白广泛存在于肝脏和脾脏组织中，被认为是肝脏疾病和癌症的血清生物标志物。铁蛋白的检测可能是健康诊断的重要工具。在这项研究中，使用了 14 个未免疫的鸡脾脏来构建单链片段 (scFv) 噬菌体文库。经过四轮淘选，获得了 7 个独特的克隆。最优克隆进一步筛选并与 NanoLuc 荧光素酶 (Nluc) 结合，作为双功能免疫探针用于双发光酶免疫分析 (BLEIA)，其灵敏度是其亲本 scFv 双夹心酶联免疫分析 (ds-ELISA) 的两倍。交叉反应性分析表明，所提出的方法具有高度选择性，适用于临床检测。为了进一步验证免疫分析的性能，用所提出的方法和商业 ELISA 试剂盒检测血清样本，结果之间具有良好的相关性。这些结果表明，与 Nluc 融合的 scFv 可能是一种快速、实用可靠、高灵敏度铁蛋白检测的强大双功能工具。
