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Research Article | Online first | Published: 01 February 2024

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

Show Author's Information Liya Ye,  Xinxin Xu,  Aihua Qu,  Liqiang Liu,  Chuanlai Xu,  Hua Kuang( )
International Joint Research Laboratory for Biointerface and Biodetection, and School of Food Science and Technology, Jiangnan University, Wuxi 214122, China
Keywords:
breast cancer, lateral flow immunoassay, monoclonal antibody, human epidermal growth factor receptor 2, quantitative
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Nano detectionDiagnosis
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Ye L, Xu X, Qu A, et al. Quantitative assessment of the breast cancer marker HER2 using a gold nanoparticle-based lateral flow immunoassay. Nano Research, 2024, https://doi.org/10.1007/s12274-024-6471-2
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Abstract Full text Electronic supplementary material About this article

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.


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About this article

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

Show Author's information Liya Ye, Xinxin Xu, Aihua Qu, Liqiang Liu, Chuanlai Xu, Hua Kuang( )
International Joint Research Laboratory for Biointerface and Biodetection, and School of Food Science and Technology, Jiangnan University, Wuxi 214122, China

Abstract

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.

Keywords: breast cancer, lateral flow immunoassay, monoclonal antibody, human epidermal growth factor receptor 2, quantitative

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Publication history
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Acknowledgements

Publication history

Received: 29 November 2023
Revised: 26 December 2023
Accepted: 03 January 2024
Published: 01 February 2024

Copyright

© Tsinghua University Press 2024

Acknowledgements

Acknowledgements

This work is financially supported by the National Natural Science Foundation of China (No. 22236002), and National Key R&D Program(Nos. 2023YFF1105003 and 2022YFA1207300).

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