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Updated: Jul 18, 2026

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
A soluble 17 kDa tumour necrosis factor (TNF) mutein, TNF-SAM2, with membrane-bound TNF-like biological
Aya Yoshida1, Chie Kohchi, Hiroyuki Inagawa
1Faculty of Engineering, The University of Tokushima, Tokushima-shi, Japan.
Background:
Membrane-bound tumour necrosis factor (TNF) has a different binding affinity to two TNF receptors (TNFR1 and TNFR2) and shows different biological activities compared with TNF-alpha. We have developed a mutein, TNF-SAM2, which has modified TNF-alpha. It has higher tumour killing activity than TNF-alpha and fewer side effects.
Materials And Methods:
The difference between TNF-SAM2 and TNF-alpha was examined in view of the binding affinity to each TNF receptor, cytotoxicity and expression of genes using Colo205 cells.
Results:
The KD value of TNF-alpha (2. 9x1-7 M) for TNFR1 was lower than TNF-SAM2 (6.9x10(-7) M). Cytotoxicity of TNF-SAM2 was significantly higher than for TNF-a. RNA of Colo205 cells stimulated with TNF-SAM2 indicated that 80 genes increased by a factor of 2 or more, and 69 genes decreased.
Conclusion:
TNF-SAM2 can induce different intracellular signal transduction in Colo205 cells compared with TNF-alpha. TNF-SAM2 may have characteristics of the membrane-bound form despite the fact that is in a soluble form.
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