Size and macromolecule stabilizer–dependent performance of gold colloids in immuno-PCR

Abstract

Gold nanoparticles (GNPs) are well-documented for their size and surface chemistry-dependent electronic and optical properties that are extensively utilized to develop highly sensitive immunoassays. GNP-based immuno-polymerase chain reaction (immuno-PCR) is especially interesting due to the facile loading of biomolecules on the surface of GNP probes and has been utilized to develop analyte-specific assays. In this study, the role of size and surface chemistry of GNPs is explored in detail to develop a highly sensitive and reproducible immuno-PCR assay for specific detection of biotinylated analytes. Our results indicate that smaller-sized gold nanoparticles outperform the larger ones in terms of their sensitivity in immuno-PCR assay and show superior loading of proteins and oligonucleotides on the surface of nanoparticles. Furthermore, the role of different macromolecular stabilizers (such as polyethylene glycol (PEG), bovine serum albumin (BSA), and PEGylated BSA) was compared to optimize the loading of biomolecules and to improve the signal-to-noise ratio of GNP probes. mPEG-BSAfunctionalized GNP probes of 15 nm were found to be highly sensitive at low concentrations of analytes and significantly (~ 30 fold) improve the limit of detection of analytes in comparison with ELISA assay.

The Integration of Gold Nanoparticles with Polymerase Chain Reaction for Constructing Colorimetric Sensing Platforms for Detection of Health-Related DNA and Proteins

“…Currently, the combination of PCR and AuNPs for the colorimetric detection of proteins is mainly limited to the step of transforming protein information into DNA information. Although AuNPs have also been explored for combination with immuno-PCR, the use of antibodies complicates the sensing systems and raise the cost of the assays, limiting their applications [ 98 , 99 , 100 ]. The discovery of aptamer renders this transformation easier and more effective [ 101 ].…”