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Matrix effect in tumor lysates - Does it affect your cytokine ELISA and multiplex analyses?
Camilla Stavnsbjerg1, Jennifer Solgaard Jørgensen1, Trine Bjørnbo Engel1
1Department of Health Technology, Section for Biotherapeutic Engineering and Drug Targeting, Technical University of Denmark, Kongens Lyngby, Denmark.
Journal of Immunological Methods
|November 10, 2021
Summary
Matrix effects significantly impact cytokine quantification in cancerous tissues using immunoassays. Careful sample preparation and dilution can mitigate but not eliminate these measurement interferences.
Area of Science:
- Oncology
- Immunology
- Biochemistry
Background:
- Cytokine quantification in cancerous tissue is crucial for tumor biology and anti-cancer drug development.
- Protein-level cytokine measurements commonly utilize immunoassays like enzyme-linked immunosorbent assays (ELISAs) and multiplex assays.
- Immunoassays are susceptible to interference from various factors, collectively termed the matrix effect, leading to inaccurate concentration readings.
Purpose of the Study:
- To investigate the presence and impact of matrix effects on cytokine measurements in tumor lysates.
- To assess the effectiveness of sample dilution and lysis buffer composition in reducing matrix effects.
Main Methods:
- Analysis of tumor lysates from 11 different syngeneic murine tumor models.
- Evaluation of cytokine measurements using ELISAs and multiplex assays.
- Assessment of the influence of sample dilution and lysis buffer components on assay interference.
Main Results:
- Matrix effects were confirmed in tumor lysates across all 11 tested murine tumor models.
- These matrix effects significantly influenced cytokine concentration measurements obtained from both ELISAs and multiplex assays.
- While dilution and buffer optimization showed some reduction, matrix effects persisted in the analyzed samples.
Conclusions:
- Matrix effects pose a significant challenge for accurate cytokine quantification in tumor lysates.
- Researchers must exercise caution when interpreting cytokine data derived from tumor samples analyzed by immunoassays.
- Further strategies may be needed to fully overcome matrix effects in complex biological samples like tumor lysates.
