Related Experiment Video
Updated: Jul 5, 2026
16:07
Determination of the Relative Potency of an Anti-TNF Monoclonal Antibody (mAb) by Neutralizing TNF Using an In Vitro Bioanalytical Method
Published on: September 16, 2017
Measurement of tumor necrosis factor alpha and beta
1National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, USA.
Current Protocols in Immunology
|April 25, 2008
Summary
This study quantifies tumor necrosis factor (TNF) activity using a sensitive cell-based assay. The method accurately measures picogram levels of TNF in various biological samples.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Tumor necrosis factor (TNF)-a and TNF-b are key cytokines involved in immune responses.
- Cellular responses to TNF binding include lysis, indicating its potent biological activity.
- Quantifying TNF levels is crucial for understanding various physiological and pathological conditions.
Purpose of the Study:
- To present a reliable protocol for quantifying TNF activity.
- To establish a method for measuring TNF in diverse biological fluids.
- To characterize the sensitivity and specificity of the developed assay.
Main Methods:
- Utilized actinomycin D-treated murine L929 fibroblasts as TNF-sensitive indicator cells.
- Developed a cell-based assay to measure TNF-induced cell lysis.
- Applied the assay to quantify TNF in cell culture supernatants, serum, and cerebrospinal fluid.
Main Results:
- The assay can detect picogram concentrations of TNF-a and TNF-b.
- Successfully quantified TNF activity in human, rat, and murine samples.
- The assay demonstrated sensitivity to both TNF-a and TNF-b, but could not differentiate between the two forms.
Conclusions:
- A robust and sensitive cell-based assay for TNF quantification has been established.
- This method is valuable for measuring TNF activity in various biological matrices.
- The assay provides a means to assess total TNF-a/b activity without distinguishing between isoforms.
