Preparation of indium-115-labeled diethylenetriaminetetraacetic acid monoacetamide peptides purified by 8-hydroxyquinoline

I. Nagy, Imre Varga, F. Hudecz

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

In this communication we describe a novel procedure for the preparation and purification of diethylenetriaminepentaacetic (DTPA)-acylated and 115In3+ -labeled oligopeptides using 8-hydroxyquinoline for the removal and quantification of nonbound indium ions. First the N(α) or N(α), N(ε)-DTPA oligopeptides containing C-terminal KDEL signal motif were produced by solid-phase synthesis. For this the free carboxyl group of DTPA dianhydride was activated in situ for a short period of time yielding a major product. Reversed-phase HPLC-purified DTPA oligopeptides were labeled with 115In3+ in aqueous buffer solution at pH 3.8. For the removal as well as for the detection of uncoordinated 115In3+ ions we have utilized the 115In3+ complex-forming ability of 8-hydroxyquinoline in chloroform. Following an optimized extraction procedure the free indium ion content was measured by spectrophotometry in the organic phase. Data obtained by this method and verified by total-reflection X-ray fluorescence spectroscopy and thin-layer chromatography demonstrated that free 115In3+ could be efficiently removed and sensitively detected in the presence of DTPA oligopeptide chelator. No release of 115n3+ from its DTPA complex was observed. This method could be useful for the preparation of indium complexes of peptides and perhaps proteins containing a DTPA moiety and nonradioactive isotope ligand. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)17-24
Number of pages8
JournalAnalytical Biochemistry
Volume287
Issue number1
DOIs
Publication statusPublished - Dec 1 2000

Fingerprint

Oxyquinoline
Oligopeptides
Indium
Peptides
Ions
X-Ray Emission Spectrometry
Thin layer chromatography
Solid-Phase Synthesis Techniques
Spectrophotometry
Chelating Agents
Thin Layer Chromatography
Chloroform
Isotopes
Purification
Buffers
High Pressure Liquid Chromatography
Ligands
diethylenetriaminetetraacetic acid
Communication
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Preparation of indium-115-labeled diethylenetriaminetetraacetic acid monoacetamide peptides purified by 8-hydroxyquinoline. / Nagy, I.; Varga, Imre; Hudecz, F.

In: Analytical Biochemistry, Vol. 287, No. 1, 01.12.2000, p. 17-24.

Research output: Contribution to journalArticle

@article{ec9d7e64bbfc49168150e296462b2311,
title = "Preparation of indium-115-labeled diethylenetriaminetetraacetic acid monoacetamide peptides purified by 8-hydroxyquinoline",
abstract = "In this communication we describe a novel procedure for the preparation and purification of diethylenetriaminepentaacetic (DTPA)-acylated and 115In3+ -labeled oligopeptides using 8-hydroxyquinoline for the removal and quantification of nonbound indium ions. First the N(α) or N(α), N(ε)-DTPA oligopeptides containing C-terminal KDEL signal motif were produced by solid-phase synthesis. For this the free carboxyl group of DTPA dianhydride was activated in situ for a short period of time yielding a major product. Reversed-phase HPLC-purified DTPA oligopeptides were labeled with 115In3+ in aqueous buffer solution at pH 3.8. For the removal as well as for the detection of uncoordinated 115In3+ ions we have utilized the 115In3+ complex-forming ability of 8-hydroxyquinoline in chloroform. Following an optimized extraction procedure the free indium ion content was measured by spectrophotometry in the organic phase. Data obtained by this method and verified by total-reflection X-ray fluorescence spectroscopy and thin-layer chromatography demonstrated that free 115In3+ could be efficiently removed and sensitively detected in the presence of DTPA oligopeptide chelator. No release of 115n3+ from its DTPA complex was observed. This method could be useful for the preparation of indium complexes of peptides and perhaps proteins containing a DTPA moiety and nonradioactive isotope ligand. (C) 2000 Academic Press.",
author = "I. Nagy and Imre Varga and F. Hudecz",
year = "2000",
month = "12",
day = "1",
doi = "10.1006/abio.2000.4833",
language = "English",
volume = "287",
pages = "17--24",
journal = "Analytical Biochemistry",
issn = "0003-2697",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Preparation of indium-115-labeled diethylenetriaminetetraacetic acid monoacetamide peptides purified by 8-hydroxyquinoline

AU - Nagy, I.

AU - Varga, Imre

AU - Hudecz, F.

PY - 2000/12/1

Y1 - 2000/12/1

N2 - In this communication we describe a novel procedure for the preparation and purification of diethylenetriaminepentaacetic (DTPA)-acylated and 115In3+ -labeled oligopeptides using 8-hydroxyquinoline for the removal and quantification of nonbound indium ions. First the N(α) or N(α), N(ε)-DTPA oligopeptides containing C-terminal KDEL signal motif were produced by solid-phase synthesis. For this the free carboxyl group of DTPA dianhydride was activated in situ for a short period of time yielding a major product. Reversed-phase HPLC-purified DTPA oligopeptides were labeled with 115In3+ in aqueous buffer solution at pH 3.8. For the removal as well as for the detection of uncoordinated 115In3+ ions we have utilized the 115In3+ complex-forming ability of 8-hydroxyquinoline in chloroform. Following an optimized extraction procedure the free indium ion content was measured by spectrophotometry in the organic phase. Data obtained by this method and verified by total-reflection X-ray fluorescence spectroscopy and thin-layer chromatography demonstrated that free 115In3+ could be efficiently removed and sensitively detected in the presence of DTPA oligopeptide chelator. No release of 115n3+ from its DTPA complex was observed. This method could be useful for the preparation of indium complexes of peptides and perhaps proteins containing a DTPA moiety and nonradioactive isotope ligand. (C) 2000 Academic Press.

AB - In this communication we describe a novel procedure for the preparation and purification of diethylenetriaminepentaacetic (DTPA)-acylated and 115In3+ -labeled oligopeptides using 8-hydroxyquinoline for the removal and quantification of nonbound indium ions. First the N(α) or N(α), N(ε)-DTPA oligopeptides containing C-terminal KDEL signal motif were produced by solid-phase synthesis. For this the free carboxyl group of DTPA dianhydride was activated in situ for a short period of time yielding a major product. Reversed-phase HPLC-purified DTPA oligopeptides were labeled with 115In3+ in aqueous buffer solution at pH 3.8. For the removal as well as for the detection of uncoordinated 115In3+ ions we have utilized the 115In3+ complex-forming ability of 8-hydroxyquinoline in chloroform. Following an optimized extraction procedure the free indium ion content was measured by spectrophotometry in the organic phase. Data obtained by this method and verified by total-reflection X-ray fluorescence spectroscopy and thin-layer chromatography demonstrated that free 115In3+ could be efficiently removed and sensitively detected in the presence of DTPA oligopeptide chelator. No release of 115n3+ from its DTPA complex was observed. This method could be useful for the preparation of indium complexes of peptides and perhaps proteins containing a DTPA moiety and nonradioactive isotope ligand. (C) 2000 Academic Press.

UR - http://www.scopus.com/inward/record.url?scp=0034544552&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034544552&partnerID=8YFLogxK

U2 - 10.1006/abio.2000.4833

DO - 10.1006/abio.2000.4833

M3 - Article

VL - 287

SP - 17

EP - 24

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

IS - 1

ER -