Effect of a purified somatostatin monoclonal antibody and its Fab fragments on gastrin release.
The effect of a purified somatostatin monoclonal antibody, SOMA 10, and its Fab fragments on gastrin release have been examined in the isolated perfused rat stomach. SOMA 10 was purified from ascites fluid by ammonium sulfate precipitation followed by hydroxylapatite (HAP) chromatography. This gave a preparation of greater than 90% purity, as assessed by fast affinity and size exclusion high-performance liquid chromatography (HPLC). HAP-purified SOMA 10 had a binding capacity of 760 mmol somatostatin/mol antibody, a dissociation constant of 2.2 nM, and recognized amino acid residues 5-12 of the somatostatin molecule. Fab fragments of SOMA 10 were prepared and purified by protein A affinity chromatography, and the purity assessed by HPLC and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Single passage perfusion of SOMA 10 (100 micrograms/ml) into the gastric vasculature caused a paradoxical decrease in basal gastrin release. However, recirculation of the antibody (20 micrograms/ml) caused an increase in cumulated gastrin release. Perfusion of Fab fragments of SOMA 10 (66 micrograms/ml) also increased gastrin secretion. These results support the suggestion that somatostatin exerts a tonic restraint on gastrin release and that penetration of the antibody to the site of somatostatin release is necessary for it to have an effect.
McIntosh
C H
CH
Department of Physiology, University of British Columbia, Vancouver, Canada.
Tang
C L
CL
Malcolm
A J
AJ
Ho
M
M
Kwok
Y N
YN
Brown
J C
JC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification / Q000494:pharmacology*; D002845:Chromatography; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000276:immunology; D005753:Gastric Mucosa / Q000276:immunology / Q000378:metabolism*; D005755:Gastrins / Q000378:metabolism*; D006882:Hydroxyapatites; D005779:Immunodiffusion; D007140:Immunoglobulin Fab Fragments / Q000302:isolation & purification; D066298:In Vitro Techniques; D007700:Kinetics; D008297:Male; D010477:Perfusion; D051381:Rats; D011919:Rats, Inbred Strains; D013004:Somatostatin / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005755:Gastrins; D006882:Hydroxyapatites; D007140:Immunoglobulin Fab Fragments; D013004:Somatostatin; D017886:Durapatite