Hydrazide-assisted directional antibody conjugation of gold nanoparticles to enhance immunochromatographic assay.

The analytical performance of immunochromatographic assay (ICA) is usually determined by the biological activity of antibody and gold nanoparticle conjugates (AuNP probes). However, conventional probes are constructed using the nondirectional coupling method that can cause the improper orientation of antibodies with the poor accessibility of antigen-binding sites. To address these issues, we report a site-specific directional coupling strategy to enhance the bioactivity of AuNP probes through the specific covalent binding of the aldehyde group in the Fc domain of antibodies with the hydrazide group modified on the surface of AuNPs. Through this design, the antibodies can be erected on the AuNP surface to fully expose the Fab domain and achieve the maximized functional availability. Leveraging these AuNP probes as ICA labels, we demonstrate an improved detection of the hepatitis B surface antigen with less used amount of labeled antibody (0.2 mg/pmol AuNPs), shorter reaction time (10 min), better antibody bioactivity, and higher detection sensitivity (2 ng/mL) compared with the carbodiimide method. Overall, this work provides great promise for the design and the construction of high-performance probes to enhance the detection performance of ICA sensors.