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 journalArticle

105 Citations (Scopus)

Abstract

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
Volume9
Issue number4
DOIs
StatePublished - Apr 28 2015

Fingerprint

immunoassay
Microarrays
bioinstrumentation
Biosensors
serums
Surface plasmon resonance
surface plasmon resonance
Microfluidics
Cytokines
Assays
antibodies
Nanorods
conjugation
Antibodies
Gold
nanorods
blood
Monitoring
Blood
chips

Keywords

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

ASJC Scopus subject areas

  • Materials Science(all)
  • Engineering(all)
  • Physics and Astronomy(all)

Cite this

Chen, P., Chung, M. T., McHugh, W., Nidetz, R., Li, Y., Fu, J., ... Kurabayashi, K. (2015). Multiplex serum cytokine immunoassay using nanoplasmonic biosensor microarrays. ACS Nano, 9(4), 4173-4181. https://doi.org/10.1021/acsnano.5b00396
Chen, Pengyu ; Chung, Meng Ting ; McHugh, Walker ; Nidetz, Robert ; Li, Yuwei ; Fu, Jianping ; Cornell, Timothy T. ; Shanley, Thomas P. ; Kurabayashi, Katsuo. / Multiplex serum cytokine immunoassay using nanoplasmonic biosensor microarrays. In: ACS Nano. 2015 ; Vol. 9, No. 4. pp. 4173-4181.
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Chen, P, Chung, MT, McHugh, W, Nidetz, R, Li, Y, Fu, J, Cornell, TT, Shanley, TP & Kurabayashi, K 2015, 'Multiplex serum cytokine immunoassay using nanoplasmonic biosensor microarrays', ACS Nano, vol. 9, no. 4, pp. 4173-4181. https://doi.org/10.1021/acsnano.5b00396

Multiplex serum cytokine immunoassay using nanoplasmonic biosensor microarrays. / Chen, Pengyu; Chung, Meng Ting; McHugh, Walker; Nidetz, Robert; Li, Yuwei; Fu, Jianping; Cornell, Timothy T.; Shanley, Thomas P.; Kurabayashi, Katsuo.

In: ACS Nano, Vol. 9, No. 4, 28.04.2015, p. 4173-4181.

Research output: Contribution to journalArticle

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Chen P, Chung MT, McHugh W, Nidetz R, Li Y, Fu J et al. Multiplex serum cytokine immunoassay using nanoplasmonic biosensor microarrays. ACS Nano. 2015 Apr 28;9(4):4173-4181. https://doi.org/10.1021/acsnano.5b00396