Direct Patterning of Antibody Conjugated Gold Nanorods Immunoassay for the Label-Free and Multiplexed Determination of Mouse Tumor-Associate Macrophages Functional Immunophenotype

Abstract

Plasmonic nanoparticle patterning has been extensively investigated over the past decades for its vast applications in miniaturized electronics, enhanced electrochemical catalysts and integrated biosensors. Current available techniques, such as electron beam lithography, dip-pen nanolithography, scanning probe microscopy-based patterning, microfluidics, and inject printing, provide multiscale capability for nanoparticle fabrication, manipulation and self-assembly patterning. However, it remains critical challenging to achieve uniform patterning via a low cost template-free method with high throughput and scalability. Herein, we report a facile method to direct pattern antibody conjugated gold nanorods (AuNRs) on a (3-Aminopropyl)triethoxysilane functionalized glass substrate in one-step manner by employing the anisotropic microstructure of weasel hairs. We successfully fabricated parallel uniform barcode nanoparticle ensembles with fine controlled widths on glass substrates over a 5cm×2cm area within less than three minutes using minimum of 3 uL AuNR solution. The fabricated device was applied as real-time and multiplexed immunoassays for functional immunophenotyping of mouse tumor-associate macrophages (TAMs) in response to BLZ in a tumor microenvironment. This nanoparticle patterning technique can be further extended to other clinical applications, such as disease diagnosis and immune status monitoring.