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In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
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Updated: Jan 19, 2026

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Reliable Sensing Platform for Plasmonic Enzyme-Linked Immunosorbent Assays Based on Automatic Flow-Based Methodology.

Natcha Kaewwonglom1, Miquel Oliver2, David J Cocovi-Solberg2

  • 1Research Center on Chemistry for Development of Health Promoting Products from Northern Resources, Department of Chemistry, Faculty of Science , Chiang Mai University , Chiang Mai 50200 , Thailand.

Analytical Chemistry
|September 10, 2019
PubMed
Summary

This study introduces an automated flow method for plasmonic enzyme-linked immunosorbent assays (ELISA), enabling ultrasensitive detection of contaminants like diclofenac. The new system offers improved reliability and lower detection limits compared to traditional batch methods.

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Area of Science:

  • Analytical Chemistry
  • Nanotechnology
  • Environmental Science

Background:

  • Plasmonic enzyme-linked immunosorbent assays (ELISA) offer ultrasensitive detection but suffer from poor ruggedness in batchwise formats.
  • Conventional ELISA methods struggle with detecting trace contaminants in complex matrices like seawater.

Purpose of the Study:

  • To develop a robust and automated flow methodology for plasmonic ELISA.
  • To enable ultrasensitive detection of small molecules and emerging contaminants using hydrogen peroxide-related enzymatic bioassays.
  • To improve the reliability and reduce the detection limits compared to existing methods.

Main Methods:

  • A competitive ELISA was integrated with in-line generation of gold nanoparticles (AuNPs).
  • Real-time monitoring of AuNP nucleation and growth rates using a miniaturized photometer.
  • A hybrid flow system with microsyringe pumps and a multiposition valve was employed for automated, computer-controlled assays.
  • Diclofenac in seawater was used as a proof-of-concept application.

Main Results:

  • The flow-based platform achieved an ultrasensitive detection limit of 0.001 μg L-1 for diclofenac, one order of magnitude lower than EU limits.
  • The system demonstrated reproducible timing in AuNP nucleation and growth, with unsupervised LSPR absorbance detection.
  • Repeatability and intermediate precision in seawater were <4% and <14%, significantly outperforming batchwise plasmonic ELISA (RSDs up to 30%).

Conclusions:

  • The automated flow plasmonic ELISA provides a reliable and ultrasensitive method for detecting emerging contaminants in complex samples.
  • This approach overcomes the limitations of batchwise plasmonic ELISA, offering enhanced ruggedness and precision.
  • The platform is suitable for high-throughput analysis and environmental monitoring applications.