Enzyme Labeling of Steroids by the N-Succinimidyl Ester Method. Preparation of Horseradish Peroxidase-Labeled Antigen for Use in Enzyme Immunoassay
Enzyme labeling of a steroid with horseradish peroxidase by the N-succinimidyl ester method has been investigated in comparison with that with β-galactosidase. The reaction of the activated ester of 4-hydroxytestosterone 4-hemiglutarate with horseradish peroxidase provided a labeled antigen showing...
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| Veröffentlicht in: | Chemical & pharmaceutical bulletin 1985/01/25, Vol.33(1), pp.249-255 |
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| container_title | Chemical & pharmaceutical bulletin |
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| creator | HOSODA, HIROSHI KARUBE, TOMOKO KOBAYASHI, NORIHIRO NAMBARA, TOSHIO |
| description | Enzyme labeling of a steroid with horseradish peroxidase by the N-succinimidyl ester method has been investigated in comparison with that with β-galactosidase. The reaction of the activated ester of 4-hydroxytestosterone 4-hemiglutarate with horseradish peroxidase provided a labeled antigen showing high immunoreactivity with an anti-testosterone antiserum in the enzyme immunoassay procedure. The effect of steroid/enzyme molar ratio, ranging from 2 to 60, in the labeling on the assay sensitivity was then examined. It was found that, in contrast to the case of β-galactosidase, the sensitivity of the assay using horseradish peroxidase-labeled antigen is not significantly influenced by the molar ratio. Dose-response curves with high sensitivities could be obtained by the use of these labeled antigens at an appropriate dilution of the antiserum. The active ester method proved to be useful for the preparation of enzymelabeled antigens because of its simplicity and excellent reproducibility. |
| doi_str_mv | 10.1248/cpb.33.249 |
| format | Article |
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Preparation of Horseradish Peroxidase-Labeled Antigen for Use in Enzyme Immunoassay</title><source>J-STAGE Free</source><source>MEDLINE</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><source>Free Full-Text Journals in Chemistry</source><creator>HOSODA, HIROSHI ; KARUBE, TOMOKO ; KOBAYASHI, NORIHIRO ; NAMBARA, TOSHIO</creator><creatorcontrib>HOSODA, HIROSHI ; KARUBE, TOMOKO ; KOBAYASHI, NORIHIRO ; NAMBARA, TOSHIO</creatorcontrib><description>Enzyme labeling of a steroid with horseradish peroxidase by the N-succinimidyl ester method has been investigated in comparison with that with β-galactosidase. The reaction of the activated ester of 4-hydroxytestosterone 4-hemiglutarate with horseradish peroxidase provided a labeled antigen showing high immunoreactivity with an anti-testosterone antiserum in the enzyme immunoassay procedure. The effect of steroid/enzyme molar ratio, ranging from 2 to 60, in the labeling on the assay sensitivity was then examined. It was found that, in contrast to the case of β-galactosidase, the sensitivity of the assay using horseradish peroxidase-labeled antigen is not significantly influenced by the molar ratio. Dose-response curves with high sensitivities could be obtained by the use of these labeled antigens at an appropriate dilution of the antiserum. The active ester method proved to be useful for the preparation of enzymelabeled antigens because of its simplicity and excellent reproducibility.</description><identifier>ISSN: 0009-2363</identifier><identifier>EISSN: 1347-5223</identifier><identifier>DOI: 10.1248/cpb.33.249</identifier><identifier>PMID: 3891119</identifier><identifier>CODEN: CPBTAL</identifier><language>eng</language><publisher>Tokyo: The Pharmaceutical Society of Japan</publisher><subject>Analysis ; Biological and medical sciences ; Esters ; General pharmacology ; Horseradish Peroxidase ; Immunoenzyme Techniques ; Medical sciences ; Pharmacology. 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Preparation of Horseradish Peroxidase-Labeled Antigen for Use in Enzyme Immunoassay</title><title>Chemical & pharmaceutical bulletin</title><addtitle>Chem. Pharm. Bull.</addtitle><description>Enzyme labeling of a steroid with horseradish peroxidase by the N-succinimidyl ester method has been investigated in comparison with that with β-galactosidase. The reaction of the activated ester of 4-hydroxytestosterone 4-hemiglutarate with horseradish peroxidase provided a labeled antigen showing high immunoreactivity with an anti-testosterone antiserum in the enzyme immunoassay procedure. The effect of steroid/enzyme molar ratio, ranging from 2 to 60, in the labeling on the assay sensitivity was then examined. It was found that, in contrast to the case of β-galactosidase, the sensitivity of the assay using horseradish peroxidase-labeled antigen is not significantly influenced by the molar ratio. Dose-response curves with high sensitivities could be obtained by the use of these labeled antigens at an appropriate dilution of the antiserum. The active ester method proved to be useful for the preparation of enzymelabeled antigens because of its simplicity and excellent reproducibility.</description><subject>Analysis</subject><subject>Biological and medical sciences</subject><subject>Esters</subject><subject>General pharmacology</subject><subject>Horseradish Peroxidase</subject><subject>Immunoenzyme Techniques</subject><subject>Medical sciences</subject><subject>Pharmacology. 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Preparation of Horseradish Peroxidase-Labeled Antigen for Use in Enzyme Immunoassay</title><author>HOSODA, HIROSHI ; KARUBE, TOMOKO ; KOBAYASHI, NORIHIRO ; NAMBARA, TOSHIO</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5489-e6cadbd95fceb17baa3087a3b352c9d88e267eccefb1dafb95c50883247a76673</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1985</creationdate><topic>Analysis</topic><topic>Biological and medical sciences</topic><topic>Esters</topic><topic>General pharmacology</topic><topic>Horseradish Peroxidase</topic><topic>Immunoenzyme Techniques</topic><topic>Medical sciences</topic><topic>Pharmacology. Drug treatments</topic><topic>sensitivity</topic><topic>Steroids - analysis</topic><topic>Succinimides</topic><topic>Testosterone - analysis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>HOSODA, HIROSHI</creatorcontrib><creatorcontrib>KARUBE, TOMOKO</creatorcontrib><creatorcontrib>KOBAYASHI, NORIHIRO</creatorcontrib><creatorcontrib>NAMBARA, TOSHIO</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Immunology Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Chemical & pharmaceutical bulletin</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>HOSODA, HIROSHI</au><au>KARUBE, TOMOKO</au><au>KOBAYASHI, NORIHIRO</au><au>NAMBARA, TOSHIO</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Enzyme Labeling of Steroids by the N-Succinimidyl Ester Method. Preparation of Horseradish Peroxidase-Labeled Antigen for Use in Enzyme Immunoassay</atitle><jtitle>Chemical & pharmaceutical bulletin</jtitle><addtitle>Chem. Pharm. Bull.</addtitle><date>1985</date><risdate>1985</risdate><volume>33</volume><issue>1</issue><spage>249</spage><epage>255</epage><pages>249-255</pages><issn>0009-2363</issn><eissn>1347-5223</eissn><coden>CPBTAL</coden><abstract>Enzyme labeling of a steroid with horseradish peroxidase by the N-succinimidyl ester method has been investigated in comparison with that with β-galactosidase. The reaction of the activated ester of 4-hydroxytestosterone 4-hemiglutarate with horseradish peroxidase provided a labeled antigen showing high immunoreactivity with an anti-testosterone antiserum in the enzyme immunoassay procedure. The effect of steroid/enzyme molar ratio, ranging from 2 to 60, in the labeling on the assay sensitivity was then examined. It was found that, in contrast to the case of β-galactosidase, the sensitivity of the assay using horseradish peroxidase-labeled antigen is not significantly influenced by the molar ratio. Dose-response curves with high sensitivities could be obtained by the use of these labeled antigens at an appropriate dilution of the antiserum. The active ester method proved to be useful for the preparation of enzymelabeled antigens because of its simplicity and excellent reproducibility.</abstract><cop>Tokyo</cop><pub>The Pharmaceutical Society of Japan</pub><pmid>3891119</pmid><doi>10.1248/cpb.33.249</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| source | J-STAGE Free; MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Free Full-Text Journals in Chemistry |
| subjects | Analysis Biological and medical sciences Esters General pharmacology Horseradish Peroxidase Immunoenzyme Techniques Medical sciences Pharmacology. Drug treatments sensitivity Steroids - analysis Succinimides Testosterone - analysis |
| title | Enzyme Labeling of Steroids by the N-Succinimidyl Ester Method. Preparation of Horseradish Peroxidase-Labeled Antigen for Use in Enzyme Immunoassay |
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