Simple, sensitive and specific quantification of diamine oxidase activity in complex matrices using newly discovered fluorophores derived from natural substrates
Objective To measure diamine oxidase (DAO) activity with high sensitivity in complex matrices like plasma or tissue extracts radioactive putrescine or horseradish peroxidase (HRP)/hydrogen peroxide (H 2 O 2 ) coupling must be used. The use of radioactive material should be avoided and HRP/H 2 O 2 co...
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| Veröffentlicht in: | Inflammation research 2020-09, Vol.69 (9), p.937-950 |
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| creator | Boehm, Thomas Karer, Matthias Gludovacz, Elisabeth Petroczi, Karin Resch, Marlene Schuetzenberger, Kornelia Klavins, Kristaps Borth, Nicole Jilma, Bernd |
| description | Objective
To measure diamine oxidase (DAO) activity with high sensitivity in complex matrices like plasma or tissue extracts radioactive putrescine or horseradish peroxidase (HRP)/hydrogen peroxide (H
2
O
2
) coupling must be used. The use of radioactive material should be avoided and HRP/H
2
O
2
coupling is compromised by antioxidants.
Methods and results
Condensation of ortho-aminobenzaldehyde (oABA) with delta-1-pyrroline and delta-1-piperideine, the autocyclization products of the DAO-oxidized natural substrates putrescine and cadaverine, generates new quinazoline fluorophores with absorption and excitation maxima of 430 and 460 nm, respectively, and peak emission at 620 nm. Fluorescent-based detection limits are 20–40 times lower compared to absorption measurements. This assay can be used to measure DAO activity in human plasma after spiking recombinant human (rh)DAO, in rat plasma after intravenous rhDAO administration, in pregnancy plasma and in tissue extracts of DAO wild-type and knock-out mice. Using rat plasma the correlation between rhDAO activity and ELISA data is 99%. Human and rat plasma without DAO spiking and tissue extracts from DAO knock-out mice showed stable and low fluorescence in the presence of high substrate concentrations.
Conclusions
Incubation of DAO with the natural substrates putrescine and cadaverine and oABA generates novel fluorophores increasing the detection limit compared to absorption measurements at least tenfold. This simple, sensitive and specific assay allows the non-radioactive quantification of DAO activity in complex matrices like plasma and tissue extracts without interference by antioxidants. |
| doi_str_mv | 10.1007/s00011-020-01359-5 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7394931</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2409194444</sourcerecordid><originalsourceid>FETCH-LOGICAL-c451t-6b5ca65411e8834d4b6e2c6796b680eec4dc3a1daa5a479579289d42d4c8609d3</originalsourceid><addsrcrecordid>eNp9kk1rFTEUhgdRbK3-AVcBNy4czdd8ZCNI0SoUumgFdyGTnLlNmUmmSeba-3P8pz3XWxS7MJucJM_7nhx4q-o1o-8Zpd2HTCllrKac1pSJRtXNk-qYSTwq2v94ijXloha9oEfVi5xvEO95z59XR4LLvhesO65-Xfp5meAdyRCyL34LxARH8gLWj96S29WEsq9M8TGQOBLnzewDkHjnncmIW1T5siM-EBv3ZndkNiV5C5ms2YcNCfBz2qEw27iFBI6M0xpTXK5jQsZBwrZ4meJMgilrMhPJ65BLMgXyy-rZaKYMrx72k-r7l89Xp1_r84uzb6efzmsrG1bqdmisaRvJGOBo0smhBW7bTrVD21MAK50VhjljGiM71XSK98pJ7qTtW6qcOKk-HnyXdZjBWQjYf9JL8rNJOx2N1_--BH-tN3GrO6GkEgwN3j4YpHi7Qi56xolhmkyAuGbNJVVMSVyIvnmE3sQ1BRwPKa5EQ1u-p_iBsinmnGD88xlG9T4B-pAAjQnQvxOgGxSJgygjHDaQ_lr_R3UPFRK3lw</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2429350624</pqid></control><display><type>article</type><title>Simple, sensitive and specific quantification of diamine oxidase activity in complex matrices using newly discovered fluorophores derived from natural substrates</title><source>SpringerNature Journals</source><creator>Boehm, Thomas ; Karer, Matthias ; Gludovacz, Elisabeth ; Petroczi, Karin ; Resch, Marlene ; Schuetzenberger, Kornelia ; Klavins, Kristaps ; Borth, Nicole ; Jilma, Bernd</creator><creatorcontrib>Boehm, Thomas ; Karer, Matthias ; Gludovacz, Elisabeth ; Petroczi, Karin ; Resch, Marlene ; Schuetzenberger, Kornelia ; Klavins, Kristaps ; Borth, Nicole ; Jilma, Bernd</creatorcontrib><description>Objective
To measure diamine oxidase (DAO) activity with high sensitivity in complex matrices like plasma or tissue extracts radioactive putrescine or horseradish peroxidase (HRP)/hydrogen peroxide (H
2
O
2
) coupling must be used. The use of radioactive material should be avoided and HRP/H
2
O
2
coupling is compromised by antioxidants.
Methods and results
Condensation of ortho-aminobenzaldehyde (oABA) with delta-1-pyrroline and delta-1-piperideine, the autocyclization products of the DAO-oxidized natural substrates putrescine and cadaverine, generates new quinazoline fluorophores with absorption and excitation maxima of 430 and 460 nm, respectively, and peak emission at 620 nm. Fluorescent-based detection limits are 20–40 times lower compared to absorption measurements. This assay can be used to measure DAO activity in human plasma after spiking recombinant human (rh)DAO, in rat plasma after intravenous rhDAO administration, in pregnancy plasma and in tissue extracts of DAO wild-type and knock-out mice. Using rat plasma the correlation between rhDAO activity and ELISA data is 99%. Human and rat plasma without DAO spiking and tissue extracts from DAO knock-out mice showed stable and low fluorescence in the presence of high substrate concentrations.
Conclusions
Incubation of DAO with the natural substrates putrescine and cadaverine and oABA generates novel fluorophores increasing the detection limit compared to absorption measurements at least tenfold. This simple, sensitive and specific assay allows the non-radioactive quantification of DAO activity in complex matrices like plasma and tissue extracts without interference by antioxidants.</description><identifier>ISSN: 1023-3830</identifier><identifier>EISSN: 1420-908X</identifier><identifier>DOI: 10.1007/s00011-020-01359-5</identifier><identifier>PMID: 32488317</identifier><language>eng</language><publisher>Cham: Springer International Publishing</publisher><subject>Absorption ; Allergology ; Antioxidants ; Biomedical and Life Sciences ; Biomedicine ; Blood plasma ; Cadaverine ; Chemical compounds ; Condensates ; Coupling ; DELTA protein ; Dermatology ; Detection limits ; Diamines ; Enzyme-linked immunosorbent assay ; Fluorescence ; Fluorophores ; Horseradish peroxidase ; Hydrogen peroxide ; Immunology ; Intravenous administration ; Neurology ; Original ; Original Research Article ; Oxidase ; Peroxidase ; Pharmacology/Toxicology ; Plasma ; Pregnancy ; Putrescine ; Radioactive materials ; Rheumatology ; Spiking ; Substrates ; Tissues</subject><ispartof>Inflammation research, 2020-09, Vol.69 (9), p.937-950</ispartof><rights>The Author(s) 2020</rights><rights>The Author(s) 2020. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c451t-6b5ca65411e8834d4b6e2c6796b680eec4dc3a1daa5a479579289d42d4c8609d3</citedby><cites>FETCH-LOGICAL-c451t-6b5ca65411e8834d4b6e2c6796b680eec4dc3a1daa5a479579289d42d4c8609d3</cites><orcidid>0000-0002-8294-0797</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00011-020-01359-5$$EPDF$$P50$$Gspringer$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00011-020-01359-5$$EHTML$$P50$$Gspringer$$Hfree_for_read</linktohtml><link.rule.ids>230,314,780,784,885,27924,27925,41488,42557,51319</link.rule.ids></links><search><creatorcontrib>Boehm, Thomas</creatorcontrib><creatorcontrib>Karer, Matthias</creatorcontrib><creatorcontrib>Gludovacz, Elisabeth</creatorcontrib><creatorcontrib>Petroczi, Karin</creatorcontrib><creatorcontrib>Resch, Marlene</creatorcontrib><creatorcontrib>Schuetzenberger, Kornelia</creatorcontrib><creatorcontrib>Klavins, Kristaps</creatorcontrib><creatorcontrib>Borth, Nicole</creatorcontrib><creatorcontrib>Jilma, Bernd</creatorcontrib><title>Simple, sensitive and specific quantification of diamine oxidase activity in complex matrices using newly discovered fluorophores derived from natural substrates</title><title>Inflammation research</title><addtitle>Inflamm. Res</addtitle><description>Objective
To measure diamine oxidase (DAO) activity with high sensitivity in complex matrices like plasma or tissue extracts radioactive putrescine or horseradish peroxidase (HRP)/hydrogen peroxide (H
2
O
2
) coupling must be used. The use of radioactive material should be avoided and HRP/H
2
O
2
coupling is compromised by antioxidants.
Methods and results
Condensation of ortho-aminobenzaldehyde (oABA) with delta-1-pyrroline and delta-1-piperideine, the autocyclization products of the DAO-oxidized natural substrates putrescine and cadaverine, generates new quinazoline fluorophores with absorption and excitation maxima of 430 and 460 nm, respectively, and peak emission at 620 nm. Fluorescent-based detection limits are 20–40 times lower compared to absorption measurements. This assay can be used to measure DAO activity in human plasma after spiking recombinant human (rh)DAO, in rat plasma after intravenous rhDAO administration, in pregnancy plasma and in tissue extracts of DAO wild-type and knock-out mice. Using rat plasma the correlation between rhDAO activity and ELISA data is 99%. Human and rat plasma without DAO spiking and tissue extracts from DAO knock-out mice showed stable and low fluorescence in the presence of high substrate concentrations.
Conclusions
Incubation of DAO with the natural substrates putrescine and cadaverine and oABA generates novel fluorophores increasing the detection limit compared to absorption measurements at least tenfold. This simple, sensitive and specific assay allows the non-radioactive quantification of DAO activity in complex matrices like plasma and tissue extracts without interference by antioxidants.</description><subject>Absorption</subject><subject>Allergology</subject><subject>Antioxidants</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Blood plasma</subject><subject>Cadaverine</subject><subject>Chemical compounds</subject><subject>Condensates</subject><subject>Coupling</subject><subject>DELTA protein</subject><subject>Dermatology</subject><subject>Detection limits</subject><subject>Diamines</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Fluorescence</subject><subject>Fluorophores</subject><subject>Horseradish peroxidase</subject><subject>Hydrogen peroxide</subject><subject>Immunology</subject><subject>Intravenous administration</subject><subject>Neurology</subject><subject>Original</subject><subject>Original Research Article</subject><subject>Oxidase</subject><subject>Peroxidase</subject><subject>Pharmacology/Toxicology</subject><subject>Plasma</subject><subject>Pregnancy</subject><subject>Putrescine</subject><subject>Radioactive materials</subject><subject>Rheumatology</subject><subject>Spiking</subject><subject>Substrates</subject><subject>Tissues</subject><issn>1023-3830</issn><issn>1420-908X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><recordid>eNp9kk1rFTEUhgdRbK3-AVcBNy4czdd8ZCNI0SoUumgFdyGTnLlNmUmmSeba-3P8pz3XWxS7MJucJM_7nhx4q-o1o-8Zpd2HTCllrKac1pSJRtXNk-qYSTwq2v94ijXloha9oEfVi5xvEO95z59XR4LLvhesO65-Xfp5meAdyRCyL34LxARH8gLWj96S29WEsq9M8TGQOBLnzewDkHjnncmIW1T5siM-EBv3ZndkNiV5C5ms2YcNCfBz2qEw27iFBI6M0xpTXK5jQsZBwrZ4meJMgilrMhPJ65BLMgXyy-rZaKYMrx72k-r7l89Xp1_r84uzb6efzmsrG1bqdmisaRvJGOBo0smhBW7bTrVD21MAK50VhjljGiM71XSK98pJ7qTtW6qcOKk-HnyXdZjBWQjYf9JL8rNJOx2N1_--BH-tN3GrO6GkEgwN3j4YpHi7Qi56xolhmkyAuGbNJVVMSVyIvnmE3sQ1BRwPKa5EQ1u-p_iBsinmnGD88xlG9T4B-pAAjQnQvxOgGxSJgygjHDaQ_lr_R3UPFRK3lw</recordid><startdate>20200901</startdate><enddate>20200901</enddate><creator>Boehm, Thomas</creator><creator>Karer, Matthias</creator><creator>Gludovacz, Elisabeth</creator><creator>Petroczi, Karin</creator><creator>Resch, Marlene</creator><creator>Schuetzenberger, Kornelia</creator><creator>Klavins, Kristaps</creator><creator>Borth, Nicole</creator><creator>Jilma, Bernd</creator><general>Springer International Publishing</general><general>Springer Nature B.V</general><scope>C6C</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7T5</scope><scope>7T7</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>BENPR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>H94</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0002-8294-0797</orcidid></search><sort><creationdate>20200901</creationdate><title>Simple, sensitive and specific quantification of diamine oxidase activity in complex matrices using newly discovered fluorophores derived from natural substrates</title><author>Boehm, Thomas ; 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Res</stitle><date>2020-09-01</date><risdate>2020</risdate><volume>69</volume><issue>9</issue><spage>937</spage><epage>950</epage><pages>937-950</pages><issn>1023-3830</issn><eissn>1420-908X</eissn><abstract>Objective
To measure diamine oxidase (DAO) activity with high sensitivity in complex matrices like plasma or tissue extracts radioactive putrescine or horseradish peroxidase (HRP)/hydrogen peroxide (H
2
O
2
) coupling must be used. The use of radioactive material should be avoided and HRP/H
2
O
2
coupling is compromised by antioxidants.
Methods and results
Condensation of ortho-aminobenzaldehyde (oABA) with delta-1-pyrroline and delta-1-piperideine, the autocyclization products of the DAO-oxidized natural substrates putrescine and cadaverine, generates new quinazoline fluorophores with absorption and excitation maxima of 430 and 460 nm, respectively, and peak emission at 620 nm. Fluorescent-based detection limits are 20–40 times lower compared to absorption measurements. This assay can be used to measure DAO activity in human plasma after spiking recombinant human (rh)DAO, in rat plasma after intravenous rhDAO administration, in pregnancy plasma and in tissue extracts of DAO wild-type and knock-out mice. Using rat plasma the correlation between rhDAO activity and ELISA data is 99%. Human and rat plasma without DAO spiking and tissue extracts from DAO knock-out mice showed stable and low fluorescence in the presence of high substrate concentrations.
Conclusions
Incubation of DAO with the natural substrates putrescine and cadaverine and oABA generates novel fluorophores increasing the detection limit compared to absorption measurements at least tenfold. This simple, sensitive and specific assay allows the non-radioactive quantification of DAO activity in complex matrices like plasma and tissue extracts without interference by antioxidants.</abstract><cop>Cham</cop><pub>Springer International Publishing</pub><pmid>32488317</pmid><doi>10.1007/s00011-020-01359-5</doi><tpages>14</tpages><orcidid>https://orcid.org/0000-0002-8294-0797</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1023-3830 |
| ispartof | Inflammation research, 2020-09, Vol.69 (9), p.937-950 |
| issn | 1023-3830 1420-908X |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7394931 |
| source | SpringerNature Journals |
| subjects | Absorption Allergology Antioxidants Biomedical and Life Sciences Biomedicine Blood plasma Cadaverine Chemical compounds Condensates Coupling DELTA protein Dermatology Detection limits Diamines Enzyme-linked immunosorbent assay Fluorescence Fluorophores Horseradish peroxidase Hydrogen peroxide Immunology Intravenous administration Neurology Original Original Research Article Oxidase Peroxidase Pharmacology/Toxicology Plasma Pregnancy Putrescine Radioactive materials Rheumatology Spiking Substrates Tissues |
| title | Simple, sensitive and specific quantification of diamine oxidase activity in complex matrices using newly discovered fluorophores derived from natural substrates |
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