Cancer Immunology, Immunotherapy

, Volume 56, Issue 4, pp 545–553

Synthesis of glycosylated human tumor necrosis factor α coupled with N-acetylneuraminic acid

  • Akiko Hayashi
  • Taku Chiba
  • Hidetoshi Hayashi
  • Satoshi Sasayama
  • Toshiyuki Ishiguro
  • Kikuo Onozaki
Original Article

DOI: 10.1007/s00262-006-0209-8

Cite this article as:
Hayashi, A., Chiba, T., Hayashi, H. et al. Cancer Immunol Immunother (2007) 56: 545. doi:10.1007/s00262-006-0209-8

Abstract

In order to study the effect of glycosylation on its biological activities, and to develop TNFα with less deleterious effects, recombinant human TNFα was chemically coupled with N-acetylneuraminic acid (NeuAc). NeuAc with C9 spacer was coupled to TNFα by acyl azide method. Two glycosylated TNFαs, designated L NeuAc-TNFα and H NeuAc-TNFα, were purified by anion-exchange chromatography. NeuAc coupling to TNFα was confirmed by lectin blotting. Average number of carbohydrate molecules introduced per molecule of L NeuAc-TNFα and H NeuAc-TNFα were estimated to be 1.0 and 1.5, respectively. We examined a variety of TNFα activities in vitro, including antiproliferative or cytotoxic activities to tumor cells, proliferative effect on fibroblast cells, stimulatory effects on IL-6 production by melanoma cells and NF-κB activation in hepatoma cells. L NeuAc-TNFα and H NeuAc-TNFα exhibited reduced activities about 1/3 and 1/10 as compared to native TNFα in all the activities performed in vitro.

Keywords

Neoglycoprotein Sialic acid Tumor necrosis factor Cytokine 

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Akiko Hayashi
    • 1
  • Taku Chiba
    • 1
  • Hidetoshi Hayashi
    • 1
  • Satoshi Sasayama
    • 1
  • Toshiyuki Ishiguro
    • 1
  • Kikuo Onozaki
    • 1
  1. 1.Department of Molecular Health Sciences, Graduate School of Pharmaceutical SciencesNagoya City University NagoyaJapan

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