Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme

<p<Abstract</p< <p<Background</p< <p<Insulin-degrading enzyme (IDE) is a widely studied zinc-metalloprotease implicated in the pathogenesis of type 2 diabetes mellitus, Alzheimer disease (AD) and varicella zoster virus infection. Despite more than six decades of researc...
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Autor*in:	Dickson Dennis W [verfasserIn] Zhao Ji [verfasserIn] Li Lilin [verfasserIn] Sahara Tomoko [verfasserIn] Reinstatler Lael [verfasserIn] Kouri Naomi [verfasserIn] DelleDonne Anthony [verfasserIn] Ertekin-Taner Nilufer [verfasserIn] Leissring Malcolm A [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2009

Übergeordnetes Werk:	In: Molecular Neurodegeneration - BMC, 2007, 4(2009), 1, p 39
Übergeordnetes Werk:	volume:4 ; year:2009 ; number:1, p 39

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.1186/1750-1326-4-39

Katalog-ID:	DOAJ054365139

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allfields	10.1186/1750-1326-4-39 doi (DE-627)DOAJ054365139 (DE-599)DOAJ5a2f3b4981ea488d84b5ad583024d94b DE-627 ger DE-627 rakwb eng RC346-429 RC952-954.6 Dickson Dennis W verfasserin aut Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme 2009 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier <p<Abstract</p< <p<Background</p< <p<Insulin-degrading enzyme (IDE) is a widely studied zinc-metalloprotease implicated in the pathogenesis of type 2 diabetes mellitus, Alzheimer disease (AD) and varicella zoster virus infection. Despite more than six decades of research on IDE, progress has been hampered by the lack of well-characterized reagents targeting this biomedically important protease. To address this important need, we generated and characterized new mouse monoclonal antibodies (mAbs) targeting natively folded human and rodent IDE.</p< <p<Results</p< <p<Eight monoclonal hybridoma cell lines were derived in house from mice immunized with full-length, natively folded, recombinant human IDE. The mAbs derived from these lines were shown to detect IDE selectively and sensitively by a wide range of methods. Two mAbs in particular—designated 6A1 and 6H9—proved especially selective for IDE in immunocytochemical and immunohistochemical applications. Using a variety of methods, we show that 6A1 selectively detects both human and rodent IDE, while 6H9 selectively detects human, but not rodent, IDE, with both mAbs showing essentially no cross reactivity with other proteins in these applications. Using these novel anti-IDE mAbs, we also developed sensitive and quantitative sandwich ELISAs capable of quantifying IDE levels present in human brain extracts.</p< <p<Conclusion</p< <p<We succeeded in developing novel mAbs that selectively detect rodent and/or human IDE, which we have shown to be suitable for a wide range of applications, including western blotting, immunoprecipitation, immunocytochemistry, immunohistochemistry, and quantitative sandwich ELISAs. These novel anti-IDE mAbs and the assays derived from them constitute important new tools for addressing many unresolved questions about the basic biology of IDE and its role in multiple highly prevalent human diseases.</p< Neurology. Diseases of the nervous system Geriatrics Zhao Ji verfasserin aut Li Lilin verfasserin aut Sahara Tomoko verfasserin aut Reinstatler Lael verfasserin aut Kouri Naomi verfasserin aut DelleDonne Anthony verfasserin aut Ertekin-Taner Nilufer verfasserin aut Leissring Malcolm A verfasserin aut In Molecular Neurodegeneration BMC, 2007 4(2009), 1, p 39 (DE-627)515978361 (DE-600)2244557-2 17501326 nnns volume:4 year:2009 number:1, p 39 https://doi.org/10.1186/1750-1326-4-39 kostenfrei https://doaj.org/article/5a2f3b4981ea488d84b5ad583024d94b kostenfrei http://www.molecularneurodegeneration.com/content/4/1/39 kostenfrei https://doaj.org/toc/1750-1326 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2009 1, p 39
spelling	10.1186/1750-1326-4-39 doi (DE-627)DOAJ054365139 (DE-599)DOAJ5a2f3b4981ea488d84b5ad583024d94b DE-627 ger DE-627 rakwb eng RC346-429 RC952-954.6 Dickson Dennis W verfasserin aut Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme 2009 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier <p<Abstract</p< <p<Background</p< <p<Insulin-degrading enzyme (IDE) is a widely studied zinc-metalloprotease implicated in the pathogenesis of type 2 diabetes mellitus, Alzheimer disease (AD) and varicella zoster virus infection. Despite more than six decades of research on IDE, progress has been hampered by the lack of well-characterized reagents targeting this biomedically important protease. To address this important need, we generated and characterized new mouse monoclonal antibodies (mAbs) targeting natively folded human and rodent IDE.</p< <p<Results</p< <p<Eight monoclonal hybridoma cell lines were derived in house from mice immunized with full-length, natively folded, recombinant human IDE. The mAbs derived from these lines were shown to detect IDE selectively and sensitively by a wide range of methods. Two mAbs in particular—designated 6A1 and 6H9—proved especially selective for IDE in immunocytochemical and immunohistochemical applications. Using a variety of methods, we show that 6A1 selectively detects both human and rodent IDE, while 6H9 selectively detects human, but not rodent, IDE, with both mAbs showing essentially no cross reactivity with other proteins in these applications. Using these novel anti-IDE mAbs, we also developed sensitive and quantitative sandwich ELISAs capable of quantifying IDE levels present in human brain extracts.</p< <p<Conclusion</p< <p<We succeeded in developing novel mAbs that selectively detect rodent and/or human IDE, which we have shown to be suitable for a wide range of applications, including western blotting, immunoprecipitation, immunocytochemistry, immunohistochemistry, and quantitative sandwich ELISAs. These novel anti-IDE mAbs and the assays derived from them constitute important new tools for addressing many unresolved questions about the basic biology of IDE and its role in multiple highly prevalent human diseases.</p< Neurology. Diseases of the nervous system Geriatrics Zhao Ji verfasserin aut Li Lilin verfasserin aut Sahara Tomoko verfasserin aut Reinstatler Lael verfasserin aut Kouri Naomi verfasserin aut DelleDonne Anthony verfasserin aut Ertekin-Taner Nilufer verfasserin aut Leissring Malcolm A verfasserin aut In Molecular Neurodegeneration BMC, 2007 4(2009), 1, p 39 (DE-627)515978361 (DE-600)2244557-2 17501326 nnns volume:4 year:2009 number:1, p 39 https://doi.org/10.1186/1750-1326-4-39 kostenfrei https://doaj.org/article/5a2f3b4981ea488d84b5ad583024d94b kostenfrei http://www.molecularneurodegeneration.com/content/4/1/39 kostenfrei https://doaj.org/toc/1750-1326 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2009 1, p 39
allfields_unstemmed	10.1186/1750-1326-4-39 doi (DE-627)DOAJ054365139 (DE-599)DOAJ5a2f3b4981ea488d84b5ad583024d94b DE-627 ger DE-627 rakwb eng RC346-429 RC952-954.6 Dickson Dennis W verfasserin aut Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme 2009 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier <p<Abstract</p< <p<Background</p< <p<Insulin-degrading enzyme (IDE) is a widely studied zinc-metalloprotease implicated in the pathogenesis of type 2 diabetes mellitus, Alzheimer disease (AD) and varicella zoster virus infection. Despite more than six decades of research on IDE, progress has been hampered by the lack of well-characterized reagents targeting this biomedically important protease. To address this important need, we generated and characterized new mouse monoclonal antibodies (mAbs) targeting natively folded human and rodent IDE.</p< <p<Results</p< <p<Eight monoclonal hybridoma cell lines were derived in house from mice immunized with full-length, natively folded, recombinant human IDE. The mAbs derived from these lines were shown to detect IDE selectively and sensitively by a wide range of methods. Two mAbs in particular—designated 6A1 and 6H9—proved especially selective for IDE in immunocytochemical and immunohistochemical applications. Using a variety of methods, we show that 6A1 selectively detects both human and rodent IDE, while 6H9 selectively detects human, but not rodent, IDE, with both mAbs showing essentially no cross reactivity with other proteins in these applications. Using these novel anti-IDE mAbs, we also developed sensitive and quantitative sandwich ELISAs capable of quantifying IDE levels present in human brain extracts.</p< <p<Conclusion</p< <p<We succeeded in developing novel mAbs that selectively detect rodent and/or human IDE, which we have shown to be suitable for a wide range of applications, including western blotting, immunoprecipitation, immunocytochemistry, immunohistochemistry, and quantitative sandwich ELISAs. These novel anti-IDE mAbs and the assays derived from them constitute important new tools for addressing many unresolved questions about the basic biology of IDE and its role in multiple highly prevalent human diseases.</p< Neurology. Diseases of the nervous system Geriatrics Zhao Ji verfasserin aut Li Lilin verfasserin aut Sahara Tomoko verfasserin aut Reinstatler Lael verfasserin aut Kouri Naomi verfasserin aut DelleDonne Anthony verfasserin aut Ertekin-Taner Nilufer verfasserin aut Leissring Malcolm A verfasserin aut In Molecular Neurodegeneration BMC, 2007 4(2009), 1, p 39 (DE-627)515978361 (DE-600)2244557-2 17501326 nnns volume:4 year:2009 number:1, p 39 https://doi.org/10.1186/1750-1326-4-39 kostenfrei https://doaj.org/article/5a2f3b4981ea488d84b5ad583024d94b kostenfrei http://www.molecularneurodegeneration.com/content/4/1/39 kostenfrei https://doaj.org/toc/1750-1326 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2009 1, p 39
allfieldsGer	10.1186/1750-1326-4-39 doi (DE-627)DOAJ054365139 (DE-599)DOAJ5a2f3b4981ea488d84b5ad583024d94b DE-627 ger DE-627 rakwb eng RC346-429 RC952-954.6 Dickson Dennis W verfasserin aut Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme 2009 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier <p<Abstract</p< <p<Background</p< <p<Insulin-degrading enzyme (IDE) is a widely studied zinc-metalloprotease implicated in the pathogenesis of type 2 diabetes mellitus, Alzheimer disease (AD) and varicella zoster virus infection. Despite more than six decades of research on IDE, progress has been hampered by the lack of well-characterized reagents targeting this biomedically important protease. To address this important need, we generated and characterized new mouse monoclonal antibodies (mAbs) targeting natively folded human and rodent IDE.</p< <p<Results</p< <p<Eight monoclonal hybridoma cell lines were derived in house from mice immunized with full-length, natively folded, recombinant human IDE. The mAbs derived from these lines were shown to detect IDE selectively and sensitively by a wide range of methods. Two mAbs in particular—designated 6A1 and 6H9—proved especially selective for IDE in immunocytochemical and immunohistochemical applications. Using a variety of methods, we show that 6A1 selectively detects both human and rodent IDE, while 6H9 selectively detects human, but not rodent, IDE, with both mAbs showing essentially no cross reactivity with other proteins in these applications. Using these novel anti-IDE mAbs, we also developed sensitive and quantitative sandwich ELISAs capable of quantifying IDE levels present in human brain extracts.</p< <p<Conclusion</p< <p<We succeeded in developing novel mAbs that selectively detect rodent and/or human IDE, which we have shown to be suitable for a wide range of applications, including western blotting, immunoprecipitation, immunocytochemistry, immunohistochemistry, and quantitative sandwich ELISAs. These novel anti-IDE mAbs and the assays derived from them constitute important new tools for addressing many unresolved questions about the basic biology of IDE and its role in multiple highly prevalent human diseases.</p< Neurology. Diseases of the nervous system Geriatrics Zhao Ji verfasserin aut Li Lilin verfasserin aut Sahara Tomoko verfasserin aut Reinstatler Lael verfasserin aut Kouri Naomi verfasserin aut DelleDonne Anthony verfasserin aut Ertekin-Taner Nilufer verfasserin aut Leissring Malcolm A verfasserin aut In Molecular Neurodegeneration BMC, 2007 4(2009), 1, p 39 (DE-627)515978361 (DE-600)2244557-2 17501326 nnns volume:4 year:2009 number:1, p 39 https://doi.org/10.1186/1750-1326-4-39 kostenfrei https://doaj.org/article/5a2f3b4981ea488d84b5ad583024d94b kostenfrei http://www.molecularneurodegeneration.com/content/4/1/39 kostenfrei https://doaj.org/toc/1750-1326 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2009 1, p 39
allfieldsSound	10.1186/1750-1326-4-39 doi (DE-627)DOAJ054365139 (DE-599)DOAJ5a2f3b4981ea488d84b5ad583024d94b DE-627 ger DE-627 rakwb eng RC346-429 RC952-954.6 Dickson Dennis W verfasserin aut Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme 2009 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier <p<Abstract</p< <p<Background</p< <p<Insulin-degrading enzyme (IDE) is a widely studied zinc-metalloprotease implicated in the pathogenesis of type 2 diabetes mellitus, Alzheimer disease (AD) and varicella zoster virus infection. Despite more than six decades of research on IDE, progress has been hampered by the lack of well-characterized reagents targeting this biomedically important protease. To address this important need, we generated and characterized new mouse monoclonal antibodies (mAbs) targeting natively folded human and rodent IDE.</p< <p<Results</p< <p<Eight monoclonal hybridoma cell lines were derived in house from mice immunized with full-length, natively folded, recombinant human IDE. The mAbs derived from these lines were shown to detect IDE selectively and sensitively by a wide range of methods. Two mAbs in particular—designated 6A1 and 6H9—proved especially selective for IDE in immunocytochemical and immunohistochemical applications. Using a variety of methods, we show that 6A1 selectively detects both human and rodent IDE, while 6H9 selectively detects human, but not rodent, IDE, with both mAbs showing essentially no cross reactivity with other proteins in these applications. Using these novel anti-IDE mAbs, we also developed sensitive and quantitative sandwich ELISAs capable of quantifying IDE levels present in human brain extracts.</p< <p<Conclusion</p< <p<We succeeded in developing novel mAbs that selectively detect rodent and/or human IDE, which we have shown to be suitable for a wide range of applications, including western blotting, immunoprecipitation, immunocytochemistry, immunohistochemistry, and quantitative sandwich ELISAs. These novel anti-IDE mAbs and the assays derived from them constitute important new tools for addressing many unresolved questions about the basic biology of IDE and its role in multiple highly prevalent human diseases.</p< Neurology. Diseases of the nervous system Geriatrics Zhao Ji verfasserin aut Li Lilin verfasserin aut Sahara Tomoko verfasserin aut Reinstatler Lael verfasserin aut Kouri Naomi verfasserin aut DelleDonne Anthony verfasserin aut Ertekin-Taner Nilufer verfasserin aut Leissring Malcolm A verfasserin aut In Molecular Neurodegeneration BMC, 2007 4(2009), 1, p 39 (DE-627)515978361 (DE-600)2244557-2 17501326 nnns volume:4 year:2009 number:1, p 39 https://doi.org/10.1186/1750-1326-4-39 kostenfrei https://doaj.org/article/5a2f3b4981ea488d84b5ad583024d94b kostenfrei http://www.molecularneurodegeneration.com/content/4/1/39 kostenfrei https://doaj.org/toc/1750-1326 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2009 1, p 39
language	English
source	In Molecular Neurodegeneration 4(2009), 1, p 39 volume:4 year:2009 number:1, p 39
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authorswithroles_txt_mv	Dickson Dennis W @@aut@@ Zhao Ji @@aut@@ Li Lilin @@aut@@ Sahara Tomoko @@aut@@ Reinstatler Lael @@aut@@ Kouri Naomi @@aut@@ DelleDonne Anthony @@aut@@ Ertekin-Taner Nilufer @@aut@@ Leissring Malcolm A @@aut@@
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issn	17501326
topic_title	RC346-429 RC952-954.6 Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme
topic	misc RC346-429 misc RC952-954.6 misc Neurology. Diseases of the nervous system misc Geriatrics
topic_unstemmed	misc RC346-429 misc RC952-954.6 misc Neurology. Diseases of the nervous system misc Geriatrics
topic_browse	misc RC346-429 misc RC952-954.6 misc Neurology. Diseases of the nervous system misc Geriatrics
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hierarchy_parent_title	Molecular Neurodegeneration
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title	Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme
ctrlnum	(DE-627)DOAJ054365139 (DE-599)DOAJ5a2f3b4981ea488d84b5ad583024d94b
title_full	Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme
author_sort	Dickson Dennis W
journal	Molecular Neurodegeneration
journalStr	Molecular Neurodegeneration
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author_browse	Dickson Dennis W Zhao Ji Li Lilin Sahara Tomoko Reinstatler Lael Kouri Naomi DelleDonne Anthony Ertekin-Taner Nilufer Leissring Malcolm A
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author-letter	Dickson Dennis W
doi_str_mv	10.1186/1750-1326-4-39
author2-role	verfasserin
title_sort	development of monoclonal antibodies and quantitative elisas targeting insulin-degrading enzyme
callnumber	RC346-429
title_auth	Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme
abstract	<p<Abstract</p< <p<Background</p< <p<Insulin-degrading enzyme (IDE) is a widely studied zinc-metalloprotease implicated in the pathogenesis of type 2 diabetes mellitus, Alzheimer disease (AD) and varicella zoster virus infection. Despite more than six decades of research on IDE, progress has been hampered by the lack of well-characterized reagents targeting this biomedically important protease. To address this important need, we generated and characterized new mouse monoclonal antibodies (mAbs) targeting natively folded human and rodent IDE.</p< <p<Results</p< <p<Eight monoclonal hybridoma cell lines were derived in house from mice immunized with full-length, natively folded, recombinant human IDE. The mAbs derived from these lines were shown to detect IDE selectively and sensitively by a wide range of methods. Two mAbs in particular—designated 6A1 and 6H9—proved especially selective for IDE in immunocytochemical and immunohistochemical applications. Using a variety of methods, we show that 6A1 selectively detects both human and rodent IDE, while 6H9 selectively detects human, but not rodent, IDE, with both mAbs showing essentially no cross reactivity with other proteins in these applications. Using these novel anti-IDE mAbs, we also developed sensitive and quantitative sandwich ELISAs capable of quantifying IDE levels present in human brain extracts.</p< <p<Conclusion</p< <p<We succeeded in developing novel mAbs that selectively detect rodent and/or human IDE, which we have shown to be suitable for a wide range of applications, including western blotting, immunoprecipitation, immunocytochemistry, immunohistochemistry, and quantitative sandwich ELISAs. These novel anti-IDE mAbs and the assays derived from them constitute important new tools for addressing many unresolved questions about the basic biology of IDE and its role in multiple highly prevalent human diseases.</p<
abstractGer	<p<Abstract</p< <p<Background</p< <p<Insulin-degrading enzyme (IDE) is a widely studied zinc-metalloprotease implicated in the pathogenesis of type 2 diabetes mellitus, Alzheimer disease (AD) and varicella zoster virus infection. Despite more than six decades of research on IDE, progress has been hampered by the lack of well-characterized reagents targeting this biomedically important protease. To address this important need, we generated and characterized new mouse monoclonal antibodies (mAbs) targeting natively folded human and rodent IDE.</p< <p<Results</p< <p<Eight monoclonal hybridoma cell lines were derived in house from mice immunized with full-length, natively folded, recombinant human IDE. The mAbs derived from these lines were shown to detect IDE selectively and sensitively by a wide range of methods. Two mAbs in particular—designated 6A1 and 6H9—proved especially selective for IDE in immunocytochemical and immunohistochemical applications. Using a variety of methods, we show that 6A1 selectively detects both human and rodent IDE, while 6H9 selectively detects human, but not rodent, IDE, with both mAbs showing essentially no cross reactivity with other proteins in these applications. Using these novel anti-IDE mAbs, we also developed sensitive and quantitative sandwich ELISAs capable of quantifying IDE levels present in human brain extracts.</p< <p<Conclusion</p< <p<We succeeded in developing novel mAbs that selectively detect rodent and/or human IDE, which we have shown to be suitable for a wide range of applications, including western blotting, immunoprecipitation, immunocytochemistry, immunohistochemistry, and quantitative sandwich ELISAs. These novel anti-IDE mAbs and the assays derived from them constitute important new tools for addressing many unresolved questions about the basic biology of IDE and its role in multiple highly prevalent human diseases.</p<
abstract_unstemmed	<p<Abstract</p< <p<Background</p< <p<Insulin-degrading enzyme (IDE) is a widely studied zinc-metalloprotease implicated in the pathogenesis of type 2 diabetes mellitus, Alzheimer disease (AD) and varicella zoster virus infection. Despite more than six decades of research on IDE, progress has been hampered by the lack of well-characterized reagents targeting this biomedically important protease. To address this important need, we generated and characterized new mouse monoclonal antibodies (mAbs) targeting natively folded human and rodent IDE.</p< <p<Results</p< <p<Eight monoclonal hybridoma cell lines were derived in house from mice immunized with full-length, natively folded, recombinant human IDE. The mAbs derived from these lines were shown to detect IDE selectively and sensitively by a wide range of methods. Two mAbs in particular—designated 6A1 and 6H9—proved especially selective for IDE in immunocytochemical and immunohistochemical applications. Using a variety of methods, we show that 6A1 selectively detects both human and rodent IDE, while 6H9 selectively detects human, but not rodent, IDE, with both mAbs showing essentially no cross reactivity with other proteins in these applications. Using these novel anti-IDE mAbs, we also developed sensitive and quantitative sandwich ELISAs capable of quantifying IDE levels present in human brain extracts.</p< <p<Conclusion</p< <p<We succeeded in developing novel mAbs that selectively detect rodent and/or human IDE, which we have shown to be suitable for a wide range of applications, including western blotting, immunoprecipitation, immunocytochemistry, immunohistochemistry, and quantitative sandwich ELISAs. These novel anti-IDE mAbs and the assays derived from them constitute important new tools for addressing many unresolved questions about the basic biology of IDE and its role in multiple highly prevalent human diseases.</p<
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title_short	Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme
url	https://doi.org/10.1186/1750-1326-4-39 https://doaj.org/article/5a2f3b4981ea488d84b5ad583024d94b http://www.molecularneurodegeneration.com/content/4/1/39 https://doaj.org/toc/1750-1326
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author2	Zhao Ji Li Lilin Sahara Tomoko Reinstatler Lael Kouri Naomi DelleDonne Anthony Ertekin-Taner Nilufer Leissring Malcolm A
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up_date	2024-07-03T22:45:09.843Z
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