Abstract
Soluble human tumor necrosis factor receptors (shTNFRI and shTNFRII) are antagonists of tumor necrosis factor-α (TNF-α) and are under clinical investigation as therapy for autoimmune diseases and transplant rejection. However, shTNFRI and shTNFRII are limited for clinical usage because of their short half-lives in vivo. Recombinant TNF-α receptors (infliximab and etanercept) are used in treatment of rheumatoid arthritis and Crohn’s disease but are also being tested for a number of other autoimmune diseases. Human serum albumin (HSA) has been used to construct long-acting fusion proteins. Here, we report the effect of fusion of HSA with shTNFRI and with shTNFRII on shTNFR’s neutralizing activity against TNF-α. HSA fusion proteins were separately expressed in Pichia pastoris. Purified recombinant shTNFRI-HSA, HSA-shTNFRI and HSA-shTNFRII could block the cytolytic activity of TNF-α in L929 cells, and the fusion at N-terminus of shTNFRI could result in larger degree of activity decline than that at the C-terminus. Activity of three fusion proteins was much weaker than etanercept, which demonstrated that fusion of HSA significantly influenced TNF-α neutralizing activity of shTNFRs. Compared with Fc fragment, HSA fusion technology may therefore not be an ideal strategy in development of long-acting shTNFRs protein drugs.
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Acknowledgments
The authors thanks for Dr. Xiaolin Kuai and Dr. Enling Li for their support in purification of techniques and determination of biological activity. This work was financially supported by Shanghai Key Technologies R&D Program of Biomedicine (Grant No.064319028).
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Zhan, J., Chen, Y., Yuan, HY. et al. Fusion of HSA influences TNF-α neutralizing activity of shTNFRs. Biotechnol Lett 34, 417–423 (2012). https://doi.org/10.1007/s10529-011-0793-3
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DOI: https://doi.org/10.1007/s10529-011-0793-3