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In vitro and in vivo evaluation of human tumor necrosis factor-alpha (hTNFalpha) chemically conjugated to monoclonal antibody.

Abstract
Human tumor necrosis-alpha (hTNF-alpha) was chemically conjugated to the murine anti-Ly-2.1 T cell antibody using heterobifunctional crosslinking agents SAMSA and SPDP. SDS-PAGE analysis of the affinity purified conjugate consisted mainly of 1:1 and 1:2 (Ly-2.1:TNF) complexes. Conjugated hTNF retained 50% of its cytotoxic activity by the L929 cytolytic assay, with an IC50 = 0.12 ng/ml. hTNF-Ly-2.1 was also cytotoxic to E3 cells (Ly-2.1+ve) with an IC50 = 1.7 microg/ml - 3 times more cytotoxic to these cells than non-conjugated hTNF in vitro. However in vivo hTNF-Ly-2.1 conjugates were more toxic to mice than hTNF. In vivo blood clearance studies in E3 tumor bearing CBF1 mice demonstrated that the half life of the conjugate was 2 hr, compared to 20 min for hTNF. In biodistribution studies, tumor accumulation of 3% was seen for hTNF-Ly-2.1 while for unconjugated hTNF no activity in tumor was detected 24hr post injection. A single dose of hTNF-Ly-2.1 increased the accumulation of 125I-anti-Ly-2.1 by 3 fold compared to controls. However, the antitumor effect of hTNF-Ly-2.1 on E3 cells in vivo was marginal with some tumor growth retardation at day 1-3. The results of these in vitro and in vivo studies on chemically conjugated h-TNF-MoAb will be helpful in the design of novel recombinant fusion proteins for targeting the biologic activity of TNF to tumours.
AuthorsG A Pietersz, B Toohey, I F McKenzie
JournalJournal of drug targeting (J Drug Target) Vol. 5 Issue 2 Pg. 109-20 ( 1998) ISSN: 1061-186X [Print] England
PMID9588867 (Publication Type: Journal Article)
Chemical References
  • Antibodies, Monoclonal
  • Immunoconjugates
  • Recombinant Proteins
  • Tumor Necrosis Factor-alpha
Topics
  • Animals
  • Antibodies, Monoclonal (chemistry)
  • Cell Line
  • Drug Evaluation
  • Half-Life
  • Humans
  • Immunoconjugates (administration & dosage, isolation & purification, pharmacokinetics)
  • Mice
  • Recombinant Proteins (administration & dosage, pharmacokinetics)
  • Thymoma (metabolism)
  • Thymus Neoplasms (metabolism)
  • Tissue Distribution
  • Tumor Necrosis Factor-alpha (chemistry, pharmacokinetics)

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