NanoLISA & NanoLISA-F: Elevating ELISA Performance

AcZon nanoparticles are spherical and hence have a surface area-to-volume ratio of 3r, where r is the radius. As r decreases, the surface area-to-volume ratio increases allowing the conjugation of a higher number of molecules. In NanoLisa, AcZon optimized the double conjugation of enzymes (horseradish peroxidase - HRP or alkaline phosphatase - AP) and secondary antibodies on the nanoparticle shell. The augmented surface allows for additional enzymes per antibody unit resulting in amplified signal. In addition, the intrinsic fluorescence of nanoparticles offers the possibility to detect analytes through this feature which more linearly correlates with analyte presence, likely because it is a direct assessment of protein expression than enzymatic reaction. It also provides a greater dynamic range, offering superior sensitivity independently from protein amount in the sample. Additionally, fluorescence-based methods facilitate multiplexing, allowing researchers to evaluate more than one protein at the same time. This means that scientists can avoid time-consuming stripping techniques which can damage the protein of interest. Additionally, in chemiluminescence-based techniques signal saturation is a particular drawback, as each enzyme used for detection has several binding sites for interaction with substrates. This exponential signal amplification leads to a rapid plateau.

Thanks to the NanoLisa line reagents (available for both HRP and AP and conjugated to different secondary antibodies) the researcher can choose the detection method which best fits the assay needs.