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Enzyme-Linked Immunosorbent Assay01:33

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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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Exploring Intensity Distributions and Sampling in SERS-Based Immunoassays.

Ariadne Tuckmantel Bido1, Arash Azarakhshi2, Alexandre G Brolo1,3

  • 1Department of Chemistry, University of Victoria, Victoria, British Columbia V8P 5C2, Canada.

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|December 1, 2022
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Summary
This summary is machine-generated.

This study demonstrates reliable quantification using surface-enhanced Raman scattering (SERS) immunoassays at picomolar concentrations. Careful experimental design and statistical analysis ensure robust and reproducible results for analytical chemistry applications.

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Area of Science:

  • Analytical Chemistry
  • Spectroscopy
  • Biotechnology

Background:

  • Surface-enhanced Raman scattering (SERS) is a sensitive spectroscopic technique with broad applications.
  • Current limitations in SERS include poor reproducibility and insufficient robustness for routine analytical chemistry.
  • Addressing these limitations is crucial for advancing SERS in quantitative assays.

Purpose of the Study:

  • To demonstrate reliable quantification using SERS immunoassays at low concentrations (picomolar range).
  • To develop a SERS-based immunoassay for quantifying IgG in human serum.
  • To establish robust experimental and statistical methodologies for SERS assays.

Main Methods:

  • Development of a SERS-based immunoassay for human IgG.
  • Utilized Nile Blue A as a SERS reporter, with calibration curves based on median band area at 592 cm-1.
  • Employed statistical analysis of 7200 SERS spectra and Scanning Electron Microscopy (SEM) for sensor characterization.

Main Results:

  • Achieved reliable quantification of IgG in human serum within the picomolar range (20.6-333 pM).
  • Demonstrated a low limit of detection (7.39 pM) and limit of quantification (60.30 pM) for IgG.
  • Observed lognormal distribution of SERS spectra and a correlation between SERS probes and signal intensity.

Conclusions:

  • Reliable SERS immunoassay quantification is achievable through meticulous experimental design and statistical analysis.
  • The developed IgG immunosensor shows high sensitivity and accuracy, with minimal error in quality control samples.
  • Sufficient sample size and interrogation of large, well-separated areas are essential for reproducible SERS measurements.