Quantitative assessment of the breast cancer marker HER2 using a gold nanoparticle-based lateral flow immunoassay  

作　　者：Liya Ye Xinxin Xu Aihua Qu Liqiang Liu Chuanlai Xu Hua Kuang 

机构地区：[1]International Joint Research Laboratory for Biointerface and Biodetection,and School of Food Science and Technology,Jiangnan University,Wuxi 214122,China

出　　处：《Nano Research》2024年第6期5452-5460,共9页纳米研究（英文版）

基　　金：the National Natural Science Foundation of China(No.22236002);National Key R&D Program(Nos.2023YFF1105003 and 2022YFA1207300).

摘　　要：Human epidermal growth factor receptor 2(HER2)is an important biomarker for detection and treatment of breast cancer.In this study,we developed monoclonal antibodies against the extracellular domain(ECD)of HER2 and established a rapid and accurate lateral flow immunoassay(LFIA)for use in community medical institutions.The gene sequence of human HER2-ECD was obtained from the National Center for Biotechnology Information(NCBI)to construct the expression plasmid.HER2-ECD protein expressed in HEK293F cells was used to immunize BALB/c mice.The monoclonal antibodies were produced in mouse ascites and isolated by hybridoma cell screening.Antibodies were analyzed for purity by SDS-PAGE(sodium dodecyl sulphate-polyacrylamide gel-electrophoresis)and affinity was assessed by enzyme-linked immunosorbent assay(ELISA)while subtypes were detected using the commercial kits.The HER2-ECD test strip was prepared based on the sandwich method and evaluated using a portable detection instrument.The affinity of the paired antibodies,4D8 and 8D9,both reached 1×108 L/mol.Both antibodies specifically recognized the HER2-ECD protein in serum.The limit of detection(LOD)of the gold nanoparticle(AuNP)-based LFIA was 1.7 ng/mL with a detection range of 1.7-400 ng/mL,and the performance of the HER2-ECD strip correlated well with that of a Siemens chemiluminescent immunoassay(CLIA)kit.In conclusion,the paired antibodies were successfully prepared with high affinity and specificity.The AuNP-based LFIA of HER2-ECD provides a fast and accurate method to detect the concentration of HER2-ECD in serum samples for clinical use in community medical institutions,and could contribute to determining the progress of the disease or the effectiveness of treatment.

关 键 词：human epidermal growth factor receptor 2 breast cancer monoclonal antibody QUANTITATIVE lateral flow immunoassay 

分 类 号：R730.51[医药卫生—肿瘤]