Promoter-regulated in vivo asymmetric self-assembly strategy to synthesize heterogeneous nanoparticles for signal amplification

Signal amplification is commonly used to enhance the sensitivity of biological analysis. Here, we present a strategy involving in vivo asymmetric self-assembly combined with promoter strength regulation to synthesize heterogeneous nanoparticles for signal amplification. Two expression vectors were constructed by genetically inserting, respectively, signal and binding molecules into the hepatitis B core antigen protein (HBcAg) structure. Because of differential expression of the two recombinant proteins in the presence of a strong promoter (T7) and a weak promoter (Tac-1) and spontaneous asymmetric self-assembly in vivo, heterogeneous HBcAg nanoparticles (NPs) with a high ratio of signal-bearing to target-binding molecules were obtained. These nanoparticles contained a large number of green fluorescent proteins as signal molecules and a small number of B1 immunoglobulin-binding domains from protein G for antibody binding, thus enabling sensitive immunoassays. As a proof of concept, improved sensitivity for antibody detection was achieved using the heterogeneous nanoparticle conjugated with a secondary antibody molecule.