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Title: Radioimmunoassay of 5-hydroxyindole acetic acid using an iodinated derivative

Abstract

A radioimmunoassay for the main catabolite of serotonin, 5-hydroxyindole acetic acid (5-HIAA), was developed by using specific antibodies and iodinated derivative. The synthesis of a /sup 125/I-iodinated analog was performed by coupling 5-HIAA to (125I-)glycyl-tyrosine without any contact between 5-HIAA and iodine or chloramine T. It was purified on a G25 Sephadex column and diluted in citrate buffer up to 2.5 X 10(5) cpm/ml. Antibodies were obtained by coupling 5-HIAA to human serum albumin with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and tested by equilibrium dialysis. After the third immunogen injection, the four rabbits gave antisera capable of binding 50% of iodinated 5-HIIA-glycyl-tyrosine at 1/2000 final dilution. A chemical conversion of the biological samples gives to the antigen molecules a better resemblance to the immunogen, thus conferring a 100-fold gain in specificity and sensitivity. This assay allows 5-HIAA to be determined in small amounts of tissue, blood, cerebrospinal fluid or perfusate without purification with a sensitivity threshold below 0.1 ng. Some applications in cat and rat are presented.

Authors:
;
Publication Date:
OSTI Identifier:
5579169
Resource Type:
Journal Article
Resource Relation:
Journal Name: J. Pharmacol. Exp. Ther.; (United States); Journal Volume: 217:3
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; IODINE 125; ISOTOPE APPLICATIONS; SEROTONIN; RADIOIMMUNOASSAY; ANTIBODIES; CATABOLISM; CATS; RATS; TYROSINE; AMINES; AMINO ACIDS; ANIMALS; AUTONOMIC NERVOUS SYSTEM AGENTS; AZOLES; BETA DECAY RADIOISOTOPES; CARBOXYLIC ACIDS; DAYS LIVING RADIOISOTOPES; DRUGS; ELECTRON CAPTURE RADIOISOTOPES; HETEROCYCLIC COMPOUNDS; HYDROXY ACIDS; HYDROXY COMPOUNDS; INDOLES; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPES; MAMMALS; METABOLISM; NEUROREGULATORS; NUCLEI; ODD-EVEN NUCLEI; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; PYRROLES; RADIOASSAY; RADIOISOTOPES; RADIOPROTECTIVE SUBSTANCES; RODENTS; SYMPATHOMIMETICS; TRACER TECHNIQUES; TRYPTAMINES; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Puizillout, J.J., and Delaage, M.A.. Radioimmunoassay of 5-hydroxyindole acetic acid using an iodinated derivative. United States: N. p., 1981. Web.
Puizillout, J.J., & Delaage, M.A.. Radioimmunoassay of 5-hydroxyindole acetic acid using an iodinated derivative. United States.
Puizillout, J.J., and Delaage, M.A.. 1981. "Radioimmunoassay of 5-hydroxyindole acetic acid using an iodinated derivative". United States. doi:.
@article{osti_5579169,
title = {Radioimmunoassay of 5-hydroxyindole acetic acid using an iodinated derivative},
author = {Puizillout, J.J. and Delaage, M.A.},
abstractNote = {A radioimmunoassay for the main catabolite of serotonin, 5-hydroxyindole acetic acid (5-HIAA), was developed by using specific antibodies and iodinated derivative. The synthesis of a /sup 125/I-iodinated analog was performed by coupling 5-HIAA to (125I-)glycyl-tyrosine without any contact between 5-HIAA and iodine or chloramine T. It was purified on a G25 Sephadex column and diluted in citrate buffer up to 2.5 X 10(5) cpm/ml. Antibodies were obtained by coupling 5-HIAA to human serum albumin with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and tested by equilibrium dialysis. After the third immunogen injection, the four rabbits gave antisera capable of binding 50% of iodinated 5-HIIA-glycyl-tyrosine at 1/2000 final dilution. A chemical conversion of the biological samples gives to the antigen molecules a better resemblance to the immunogen, thus conferring a 100-fold gain in specificity and sensitivity. This assay allows 5-HIAA to be determined in small amounts of tissue, blood, cerebrospinal fluid or perfusate without purification with a sensitivity threshold below 0.1 ng. Some applications in cat and rat are presented.},
doi = {},
journal = {J. Pharmacol. Exp. Ther.; (United States)},
number = ,
volume = 217:3,
place = {United States},
year = 1981,
month = 6
}
  • A major handicap in the development of simple and accurate radioimmunoassay procedures for bile acids has been the lack of a radioactive standard of high specific activity. To provide such a compound, we first synthesized cholylhistamine using the carbodiimide reaction. The hypothesized structure was confirmed by elemental analysis, thin-layer chromatography, infrared and mass spectral analysis. The cholylhistamine was then iodinated with /sup 125/I, using the choloramine-T method. The /sup 125/I-cholylhistamine was bound by antisera raised against a cholic acid-bovine serum albumin conjugate. This procedure should prove useful in preparing radioactive conjugates for all of the bile acids.
  • The ability of an iodinated fatty acid, iodine-123 phenylpentadecanoic acid (1-123 PPA), and single-photon emission computed tomography (SPECT) to detect myocardium injured by temporary or permanent coronary arterial occlusion was evaluated. In 5 control dogs, 11 dogs that underwent 90 to 120 minutes of fixed left anterior descending coronary artery (LAD) occlusion, and 8 dogs that underwent 90 minutes of temporary LAD occlusion and up to 90 minutes of reflow, 2 to 6 mCi of I-123 PPA were injected and the dogs were imaged with SPECT. Control dogs showed relatively uniform uptake and clearance of I-123 PPA in similar leftmore » ventricular (LV) regions. Dogs with permanent LAD occlusion were identified by computer algorithm as having regions of decreased I-123 PPA uptake in the infarct-related area and a reduced rate of I-123 PPA clearance (-9.4% in infarct sectors (washin), +3.7% in sectors adjacent to the area of infarction, and +15.4% in control LV sectors (p less than 0.01)). Dogs with temporary LAD occlusion and reperfusion had decreased clearance of I-123 PPA from the regions with infarction; I-123 PPA clearance was -5.2 +/- 16.4% in infarct sectors, 12.7 +/- 7.4% in periinfarct zones, and 30.4 +/- 12% in control LV regions. These data demonstrate that tomographic analysis of I-123 PPA uptake and clearance permits the relatively noninvasive detection of LV myocardium injured by permanent or temporary LAD occlusion and reperfusion.« less