Multiplex serum cytokine immunoassay using nanoplasmonic biosensor microarrays

Pengyu Chen, Meng Ting Chung, Walker McHugh, Robert Nidetz, Yuwei Li, Jianping Fu, Timothy T. Cornell, Thomas P. Shanley, Katsuo Kurabayashi*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

253 Scopus citations


Precise monitoring of the rapidly changing immune status during the course of a disease requires multiplex analysis of cytokines from frequently sampled human blood. However, the current lack of rapid, multiplex, and low volume assays makes immune monitoring for clinical decision-making (e.g., critically ill patients) impractical. Without such assays, immune monitoring is even virtually impossible for infants and neonates with infectious diseases and/or immune mediated disorders as access to their blood in large quantities is prohibited. Localized surface plasmon resonance (LSPR)-based microfluidic optical biosensing is a promising approach to fill this technical gap as it could potentially permit real-time refractometric detection of biomolecular binding on a metallic nanoparticle surface and sensor miniaturization, both leading to rapid and sample-sparing analyte analysis. Despite this promise, practical implementation of such a microfluidic assay for cytokine biomarker detection in serum samples has not been established primarily due to the limited sensitivity of LSPR biosensing. Here, we developed a high-throughput, label-free, multiarrayed LSPR optical biosensor device with 480 nanoplasmonic sensing spots in microfluidic channel arrays and demonstrated parallel multiplex immunoassays of six cytokines in a complex serum matrix on a single device chip while overcoming technical limitations. The device was fabricated using easy-to-implement, one-step microfluidic patterning and antibody conjugation of gold nanorods (AuNRs). When scanning the scattering light intensity across the microarrays of AuNR ensembles with dark-field imaging optics, our LSPR biosensing technique allowed for high-sensitivity quantitative cytokine measurements at concentrations down to 5-20 pg/mL from a 1 μL serum sample. Using the nanoplasmonic biosensor microarray device, we demonstrated the ability to monitor the inflammatory responses of infants following cardiopulmonary bypass (CPB) surgery through tracking the time-course variations of their serum cytokines. The whole parallel on-chip assays, which involved the loading, incubation, and washing of samples and reagents, and 10-fold replicated multianalyte detection for each sample using the entire biosensor arrays, were completed within 40 min.

Original languageEnglish (US)
Pages (from-to)4173-4181
Number of pages9
JournalACS nano
Issue number4
StatePublished - Apr 28 2015


  • localized surface plasmon resonance (LSPR)
  • multiplexed immunoassay
  • nanoplasmonic sensing
  • optofluidics
  • serum cytokines

ASJC Scopus subject areas

  • General Engineering
  • General Physics and Astronomy
  • General Materials Science


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