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 Citations (Scopus)

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

Fingerprint

Cross-Linking Reagents
Sodium Dodecyl Sulfate
Serum Albumin
Polyacrylamide Gel Electrophoresis
Western Blotting
human TNF protein
ethylene
Serum

ASJC Scopus subject areas

  • Cancer Research
  • Immunology
  • Pharmacology

Cite this

@article{7824927e795b4a4c8e231890d185eac0,
title = "Analysis of the molecular organization of recombinant human tumor necrosis factor (rTNF) in solution using ethylene glycolbis (succinimidylsuccinate) as the cross-linking reagent",
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.",
author = "Kit Lam and P. Scuderi and Salmon, {S. E.}",
year = "1988",
language = "English (US)",
volume = "7",
pages = "267--275",
journal = "Journal of Immunotherapy",
issn = "1524-9557",
publisher = "Lippincott Williams and Wilkins",
number = "3",

}

TY - JOUR

T1 - Analysis of the molecular organization of recombinant human tumor necrosis factor (rTNF) in solution using ethylene glycolbis (succinimidylsuccinate) as the cross-linking reagent

AU - Lam, Kit

AU - Scuderi, P.

AU - Salmon, S. E.

PY - 1988

Y1 - 1988

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0023923940&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023923940&partnerID=8YFLogxK

M3 - Article

C2 - 3392553

AN - SCOPUS:0023923940

VL - 7

SP - 267

EP - 275

JO - Journal of Immunotherapy

JF - Journal of Immunotherapy

SN - 1524-9557

IS - 3

ER -