Analysis of the molecular organization of recombinant human tumor necrosis factor (rTNF) in solution using ethylene glycolbis (succinimidylsuccinate) as the cross-linking reagent

Kit Lam, P. Scuderi, S. E. Salmon

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Analysis of chemically cross-linked recombinant human tumor necrosis factor (rTNF) on sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed the presence of three distinct molecular forms which correspond to trimers, dimers, and monomers. The cross-linking procedure appeared to enhance the antigenicity of these molecular forms of rTNF as determined by Western blot analysis. Cross-linking was unaffected by 2% Nonidet P-40, high concentrations of human serum albumin, or 60% normal human serum. Sodium dodecyl sulfate (0.08%) completely abolished the cross-linking of rTNF. This paper demonstrates that rTNF, at a concentration as low as 0.8 ng/ml, exists solely in the trimeric form in solution under nondenaturing conditions.

Original languageEnglish (US)
Pages (from-to)267-275
Number of pages9
JournalJournal of Biological Response Modifiers
Volume7
Issue number3
StatePublished - 1988
Externally publishedYes

ASJC Scopus subject areas

  • Cancer Research
  • Immunology
  • Pharmacology

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