Iodination of proteins, glycoproteins, and peptides using a solid-phase oxidizing agent, 1,3,4,6-tetrachloro-3α,6α-diphenyl glycoluril (Iodogen)

doi:10.1016/0003-2697(81)90703-X

Analytical Biochemistry

Volume 117, Issue 1, October 1981, Pages 136-146

https://doi.org/10.1016/0003-2697(81)90703-X Get rights and content

Abstract

A new, commercially available oxidizing agent, 1,3,4,6-tetrachloro-3α,6α-diphenyl glycoluril (Iodogen) was compared with chloramine-T and solid-phase lactoperoxidase in the radioiodination of proteins, glycoproteins, and peptides. A method for performing low-level iodinations is described and was used to determine maximum ¹²⁵I incorporation. Iodinated proteins were purified on analytical gel filtration columns and peptides by reverse-phase high-performance liquid chromatography. Both methods were designed to analyze the tracers for the presence of aggregate and breakdown products caused by the iodination. All tracers prepared were tested in antibody dilution and dose-response curves in their respective radioimmunoassays. Results indicate that Iodogen can be used for a wide range of proteins and peptides, can permit theoretical iodine incorporation with minimal oxidation damage, and can produce tracer stable for up to 3 months.

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Citation Excerpt :
The obtained results revealed a stoichiometry of 27.3 ± 0.4 (mean ± standard deviation, SD, n = 3) moles of iodine per mole of antibody. By consulting the antibody sequence in the UniProtKB database, it was possible to obtain that the antibody contains 26 tyrosine residues, the amino acid that is more likely iodinated under the proposed conditions, as previously reported by other authors [29]. One of the risks associated to antibody labelling with iodine refers to the loss of antibody recognition capabilities upon reaction.
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Iodination of proteins, glycoproteins, and peptides using a solid-phase oxidizing agent, 1,3,4,6-tetrachloro-3α,6α-diphenyl glycoluril (Iodogen)

Abstract

Biochim. Biophys. Acta

Biochem. Biophys. Res. Commun

Nature (London)

Biochem. J

Biochem. J

Cell. Biol

J. Endocrinol

J. Endocrinol

Biochem. Biophys. Res. Commun

J. Biol. Chem

Biochemistry