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A monoclonal antibody to somatostatin with potent in vivo immunoneutralizing activity

Authors
Journal
Peptides
0196-9781
Publisher
Elsevier
Publication Date
Volume
11
Issue
4
Identifiers
DOI: 10.1016/0196-9781(90)90185-8
Keywords
  • Somatostatin
  • Monoclonal Antibody
  • Growth Hormone Release
  • Gastric Secretion
  • Gastrin
  • Immunohistochemistry
  • Paracrine Actions
  • Immunoneutralization
Disciplines
  • Biology
  • Chemistry
  • Medicine

Abstract

Abstract The spleen from a Robertsonian mouse with high titer and affinity antiserum after being immunized with somatostatin-14 conjugated to keyhole limpet hemocyanin was fused with FOX-NY cells. Hybridomas were cloned by limiting dilution, subcloned, and ascites was produced from the highest affinity clone in pristine-primed Balb/c mice. Ascites fluid contained approximately 20 mg/ml IgG and bound 50% of 1 fmol 125I-[Tyr 1]-somatostatin at a final dilution of 1:10,000,000. Binding of this IgG 1 antibody, CURE.S6, was inhibited by 50% at 40 pM concentrations of either somatostatin-14 or somatostatin-28, but was not inhibited by [D-Trp 8]-somatostatin at 1000-fold higher concentrations. The antibody produced very intense specific immunohistochemical staining of somatostatin endocrine cells in the stomach and pancreas and of intestinal somatostatin neurons with extremely low background staining. Intravenous injection of 2 mg purified antibody in urethane-anesthetized rats resulted in 300-fold increase in plasma GH within 15 min. CURE.S6 is a high affinity monoclonal antibody directed at the biologically active somatostatin ring structure. This antibody is useful for in vivo immunoneutralization of exogenous and endogenous somatostatin in the rat and also is an excellent reagent for immunohistochemical localization of somatostatin.

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