Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

The role of cortisol in immunosuppression in subarachnoid haemorrhage.

European journal of medical research·2023
Same author

SCIL-STROKE (Subcutaneous Interleukin-1 Receptor Antagonist in Ischemic Stroke): A Randomized Controlled Phase 2 Trial.

Stroke·2018
Same author

Reduction of inflammation after administration of interleukin-1 receptor antagonist following aneurysmal subarachnoid hemorrhage: results of the Subcutaneous Interleukin-1Ra in SAH (SCIL-SAH) study.

Journal of neurosurgery·2017
Same author

Diagnosis of Stroke-Associated Pneumonia: Recommendations From the Pneumonia in Stroke Consensus Group.

Stroke·2015
Same author

How is pneumonia diagnosed in clinical stroke research? A systematic review and meta-analysis.

Stroke·2015
Same author

The effect of intravenous interleukin-1 receptor antagonist on inflammatory mediators in cerebrospinal fluid after subarachnoid haemorrhage: a phase II randomised controlled trial.

Journal of neuroinflammation·2014

Related Experiment Video

Updated: May 9, 2026

Multiplex Cytokine Profiling of Stimulated Mouse Splenocytes Using a Cytometric Bead-based Immunoassay Platform
11:00

Multiplex Cytokine Profiling of Stimulated Mouse Splenocytes Using a Cytometric Bead-based Immunoassay Platform

Published on: November 9, 2017

Overcoming matrix matching problems in multiplex cytokine assays.

Margaret E Hoadley1, Stephen J Hopkins

  • 1Brain Injury Research Group, Clinical Sciences Building, Salford Royal Foundation Trust, Salford, M6 8HD, UK.

Journal of Immunological Methods
|August 7, 2013
PubMed
Summary

Matching assay matrices to samples is crucial for accurate immunoassay results, especially in multiplex cytokine assays. Strategies using horse serum improved matrix matching, ensuring reliable analyte recovery from human plasma.

Keywords:
Assay matrixCytokinesILImmunoassayMCP-1MultiplexTNF-αinterleukinmonocyte chemoattractant protein-1tumour necrosis factor-α

More Related Videos

Using Reference Reagents to Confirm Robustness of Cytokine Release Assays for the Prediction of Monoclonal Antibody Safety
06:37

Using Reference Reagents to Confirm Robustness of Cytokine Release Assays for the Prediction of Monoclonal Antibody Safety

Published on: September 15, 2023

Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry
09:15

Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry

Published on: July 26, 2019

Related Experiment Videos

Last Updated: May 9, 2026

Multiplex Cytokine Profiling of Stimulated Mouse Splenocytes Using a Cytometric Bead-based Immunoassay Platform
11:00

Multiplex Cytokine Profiling of Stimulated Mouse Splenocytes Using a Cytometric Bead-based Immunoassay Platform

Published on: November 9, 2017

Using Reference Reagents to Confirm Robustness of Cytokine Release Assays for the Prediction of Monoclonal Antibody Safety
06:37

Using Reference Reagents to Confirm Robustness of Cytokine Release Assays for the Prediction of Monoclonal Antibody Safety

Published on: September 15, 2023

Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry
09:15

Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry

Published on: July 26, 2019

Area of Science:

  • Biochemistry
  • Immunology
  • Analytical Chemistry

Background:

  • Immunoassay accuracy depends on matching assay matrices with sample matrices.
  • Multiplex assays require a single matrix suitable for all analytes, posing significant challenges.
  • Matrix effects can lead to inaccurate quantification of analytes in biological samples.

Purpose of the Study:

  • To develop strategies for overcoming matrix effects in a human plasma cytokine multiplex assay.
  • To identify and mitigate interference from plasma components in immunoassay development.
  • To optimize assay conditions for reliable cytokine quantification in human plasma.

Main Methods:

  • Diluting standard analytes in plasma to identify representative samples for assay development.
  • Screening horse sera for potential interference before use in matrix adjustment.
  • Adjusting the assay buffer matrix using horse serum to match plasma characteristics.
  • Evaluating the recovery of known analyte concentrations from plasma to confirm matrix match suitability.

Main Results:

  • Individual human plasmas variably affected assay signals for certain analytes, notably IL-1α and IL-1β.
  • Incorporating horse serum into the assay buffer enhanced matrix matching to plasma samples.
  • Endogenous monocyte chemoattractant protein-1 (MCP-1) activity was detected in one plasma sample.
  • Successful matrix matching achieved analyte recoveries within 10%–20% of expected values in most cases.

Conclusions:

  • Careful selection of samples and diluents is critical for developing robust cytokine multiplex assays.
  • Matrix matching strategies, including the use of adjuvants like horse serum, can significantly improve assay accuracy.
  • While effective, these methods may not fully account for atypical interfering activity present in some biological samples.