Related Experiment Video
Updated: Jul 5, 2026
07:18
Highly-Multiplexed Tissue Imaging with Raman Dyes
Published on: April 21, 2022
Immunoassay for P38 MAPK using surface enhanced resonance Raman spectroscopy (SERRS)
Phil Douglas1, Robert J Stokes, Duncan Graham
1Centre of Molecular Nanometrology, Department of Pure & Applied Chemistry, University of Strathclyde, 298 Cathedral Street, Glasgow, UK.
The Analyst
|May 22, 2008
Summary
This study introduces a sensitive micro-bead assay for detecting P38 mitogen-activated protein kinase. Surface-enhanced resonance Raman spectroscopy (SERRS) enabled highly sensitive quantification of the target protein in biological samples.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Spectroscopy
Background:
- P38 mitogen-activated protein kinase (MAPK) is a crucial signaling molecule involved in cellular responses.
- Accurate and sensitive detection methods for P38 MAPK are essential for understanding cellular processes and disease mechanisms.
- Existing immunoassay techniques may require further optimization for enhanced sensitivity and quantification.
Purpose of the Study:
- To develop and characterize a novel micro-bead based sandwich immunoassay for the sensitive detection of P38 MAPK.
- To utilize Surface-Enhanced Resonance Raman Spectroscopy (SERRS) for quantitative signal detection.
- To investigate the underlying mechanism of SERRS detection within the immunoassay.
Main Methods:
- A sandwich immunoassay format was designed using magnetic micro-beads for antibody immobilization.
- Monoclonal capture antibodies and a rhodamine-labelled secondary antibody were employed.
- Surface-Enhanced Resonance Raman Spectroscopy (SERRS) was utilized for quantitative detection of the labelled antibody.
- Scanning electron microscopy (SEM) was used to analyze the SERRS signal generation mechanism.
Main Results:
- The developed assay demonstrated high sensitivity with a limit of detection of 9.5 x 10(-12) mol dm(-3) for P38 MAPK.
- The quantitative sandwich assay achieved a sensitivity of 6 ng ml(-1).
- Investigation revealed that colloidal suspensions, not adsorbed nanoparticles, generated the SERRS signals, with aggregate size and organic matter distribution influencing the results.
Conclusions:
- A highly sensitive and quantitative micro-bead sandwich immunoassay for P38 MAPK detection using SERRS has been successfully developed.
- The study elucidated the mechanism of SERRS signal generation in this immunoassay, highlighting the role of colloidal suspensions.
- This SERRS-based approach offers a promising tool for sensitive biomarker detection in biological research and diagnostics.
