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Multiplex serum cytokine immunoassay using nanoplasmonic biosensor microarrays.

Pengyu Chen1, Meng Ting Chung1, Walker McHugh2

  • 1†Department of Mechanical Engineering, University of Michigan, Ann Arbor, Michigan 48109, United States.

ACS Nano
|March 21, 2015
PubMed
Summary

This study introduces a novel localized surface plasmon resonance (LSPR) biosensor for rapid, multiplex cytokine analysis in small blood volumes. The high-sensitivity device enables precise immune status monitoring, crucial for critically ill patients and infants.

Keywords:
localized surface plasmon resonance (LSPR)multiplexed immunoassaynanoplasmonic sensingoptofluidicsserum cytokines

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Area of Science:

  • Biomedical Engineering
  • Nanotechnology
  • Immunology

Background:

  • Precise immune status monitoring requires multiplex cytokine analysis from frequent blood samples.
  • Current assays are impractical for clinical decision-making due to slow speed, low multiplexing, and large volume requirements.
  • Infant immune monitoring is particularly challenging due to limited blood volume access.

Purpose of the Study:

  • To develop a high-throughput, label-free, multiarrayed localized surface plasmon resonance (LSPR) optical biosensor for sensitive multiplex cytokine detection.
  • To overcome the limited sensitivity of existing LSPR biosensing techniques for practical implementation in complex biological matrices.
  • To enable rapid and sample-sparing immune monitoring for clinical applications, especially in vulnerable populations like infants.

Main Methods:

  • Fabrication of a microfluidic device with 480 nanoplasmonic sensing spots using microfluidic patterning and gold nanorod (AuNR) antibody conjugation.
  • Development of a high-throughput, label-free LSPR optical biosensing technique utilizing dark-field imaging to scan scattering light intensity.
  • Demonstration of parallel multiplex immunoassays for six cytokines in a complex serum matrix using only 1 μL of sample.

Main Results:

  • Achieved high-sensitivity quantitative cytokine measurements down to 5-20 pg/mL.
  • Successfully demonstrated parallel multiplex detection of six cytokines in serum on a single chip.
  • Completed whole on-chip assays, including sample loading, incubation, washing, and 10-fold replicated multianalyte detection, within 40 minutes.
  • Validated the device's utility by monitoring inflammatory responses in infants post-cardiopulmonary bypass surgery.

Conclusions:

  • The developed multiarrayed LSPR biosensor offers a rapid, sensitive, and sample-sparing solution for multiplex cytokine analysis.
  • This technology overcomes previous sensitivity limitations, enabling practical implementation for immune status monitoring in complex samples.
  • The biosensor is suitable for real-time monitoring of inflammatory responses, particularly in clinical settings involving infants and critically ill patients.