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The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Application to the radioimmunoassay

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Bolton, A. E., Hunter, W. M.·1973·Biochemical Journal
DOI 10.1042/bj1330529
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Abstract

1. A new method is described for labelling proteins to high specific radioactivities with 125I. The protein is treated with a 125I-labelled acylating agent, iodinated 3-(4-hydroxyphenyl)propionic acid N-hydroxysuccinimide ester, which reacts with free amino groups in the protein molecule to attach the 125I-labelled groups by amide bonds. 2. Three protein hormones have been labelled by this method, human growth hormone, human thyroid-stimulating hormone and human luteinizing hormone. Specific radioactivities of up to 170, 120 and 55μCi/μg respectively have been obtained for these hormones. 3. The immunoreactivity of these labelled hormones has been investigated by using a radioimmunoassay system specific for each hormone. These preparations have also been compared with and found to be equal or superior to labelled hormones prepared by chemical substitution of 125I into tyrosine residues of the proteins by using the chloramine-t-oxidation procedure. 4. With some antisera the immunoreactivity of the antigen was diminished by the introduction of a single I atom into the tyrosyl groups, whereas antigen containing a single 125I-labelled 3-(4-hydroxyphenyl)propionamide group showed the same immunoreactivity as the unmodified antigen.

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Metadata

Authors
Bolton, A. E., & Hunter, W. M.
Venue
Biochemical Journal
Publisher
Portland Press Ltd.
Published
1973-07-01
Volume
133
Issue
3
Pages
529-538
ISSN
0264-6021 (ISSN-L)
DOI
10.1042/bj1330529
OpenAlex
W258376682
Cited by
3,254
References
0
FWCI
36.59
Type
journal article
Language
en
Open access
bronze

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