Multiplexed gold nanorod array biochip for multi-sample analysis

Optical transduction of biological bindings based on localized surface plasmon resonance (LSPR) of gold nanorods (GNRs) is attractive for label-free biosensing. The aspect ratio (AR) dependence of LSPR band maxima inherently provides an ideal multiplex mechanism. GNRs of selected sizes can be combined to ensure distinct plasmon peaks in absorption spectrum. Monitoring the spectral shift at the dedicated peaks allows for simultaneous detection of the specific analyte. Here, we first transformed the GNR's multiplexed biosensing capability to a robust chip-based format. Specifically, nanorods of AR 2.6 and 4.5 were assembled onto thiol-terminated substrates, followed by functionalization of respective antibodies to construct a GNR multiplex biochip. As a model system, concentrations of human IgG and rabbit IgG were simultaneously measured by correlating red-shifts at distinct resonance peaks caused by specific target binding. The calibration curves exhibited linear relationship between the spectral shift and analyte amount. The sensing performance in multi-analyte mode correlated nicely with those for single analyte detection with minimal cross-reactivity. Moreover, mixed GNRs can be deposited in controllable array pattern on the glass chip to analyze numerous samples at the same time. Each GNRs dot functioned independently as a multiplexed plamonic sensor. Coupled with microplate reader, this GNR nanoarray chip can potentially result in large scale assay of samples concurrently while for each sample, a multi-analyte detection simultaneously if desired. The concept shown in this work is simple and versatile that will definitely be a new paradigm in high-throughput protein biochip development in the era of nano-biosensing.