Monoclonal antibodies 1B3 and 5C8 as probes for monitoring the integrity of the C-terminal end of soluble angiotensin-converting enzyme

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dc.contributor.authorBalyasnikova, I V
dc.contributor.authorSun, Z L
dc.contributor.authorBerestetskaya, Y V
dc.contributor.authorChubb, A J
dc.contributor.authorAlbrecht, R F
dc.contributor.authorSturrock, E D
dc.contributor.authorDanilov, S M
dc.date.accessioned2016-09-01T07:47:47Z
dc.date.available2016-09-01T07:47:47Z
dc.date.issued2005
dc.date.updated2016-08-30T11:05:29Z
dc.description.abstractAngiotensin-converting enzyme (ACE) is a membrane-anchored ectoprotein that is proteolytically cleaved, yielding an enzymatically active soluble ACE. Two mouse monoclonal antibodies, MAbs 1B3 and 5C8, were generated to the C-terminal part of human soluble ACE. MAb 1B3 recognized the catalytically active ACE, as revealed by ELISA and precipitation assays, whereas Western blotting and immunohistochemisty on paraffin- embedded sections using MAb 5C8 detected denatured ACE. MAb 1B3 showed extensive cross-reactivity, recognizing 15 species out of the 16 tested. The binding of this MAb to ACE was greatly affected by conformational changes induced by adsorption on plastic, formalin fixation, and underglycosylation. Furthermore, MAb 1B3 binding to the mutated ACE (Pro1199Leu substitution in the juxtamembrane region, leading to a fivefold increase in serum ACE level) was markedly decreased. MAb 5C8 detected all the known expression sites of full-size ACE using formalin-fixed and paraffin-embedded human tissues. The sequential epitope for MAb 5C8 is formed by the last 11 amino acid residues of soluble ACE (Pro1193–Arg1203), whereas the conformational epitope for 1B3 is formed by a motif within these 11 amino acid residues and, in addition, by at least one stretch that includes Ala837–His839 located distal to the sequential epitope. Our findings demonstrated that MAbs 1B3 and 5C8 are very useful for the study of ACE shedding, for identification of mutations in stalk regions, and for studying alternatively spliced variants of ACE. In addition, binding of MAb 1B3 is a sensitive determinant of the integrity of soluble ACE.en_ZA
dc.identifierhttp://dx.doi.org/10.1089/hyb.2005.24.14
dc.identifier.apacitationBalyasnikova, I. V., Sun, Z. L., Berestetskaya, Y. V., Chubb, A. J., Albrecht, R. F., Sturrock, E. D., & Danilov, S. M. (2005). Monoclonal antibodies 1B3 and 5C8 as probes for monitoring the integrity of the C-terminal end of soluble angiotensin-converting enzyme. <i>Hybridoma</i>, http://hdl.handle.net/11427/21633en_ZA
dc.identifier.chicagocitationBalyasnikova, I V, Z L Sun, Y V Berestetskaya, A J Chubb, R F Albrecht, E D Sturrock, and S M Danilov "Monoclonal antibodies 1B3 and 5C8 as probes for monitoring the integrity of the C-terminal end of soluble angiotensin-converting enzyme." <i>Hybridoma</i> (2005) http://hdl.handle.net/11427/21633en_ZA
dc.identifier.citationBalyasnikova, I. V., Sun, Z. L., Franke, F. E., Berestetskaya, Y. V., Chubb, A. J., Albrecht, R. F., ... & Danilov, S. M. (2005). Monoclonal antibodies 1B3 and 5C8 as probes for monitoring the integrity of the C-terminal end of soluble angiotensin-converting enzyme. Hybridoma, 24(1), 14-26.en_ZA
dc.identifier.issn1554-0014en_ZA
dc.identifier.ris TY - Journal Article AU - Balyasnikova, I V AU - Sun, Z L AU - Berestetskaya, Y V AU - Chubb, A J AU - Albrecht, R F AU - Sturrock, E D AU - Danilov, S M AB - Angiotensin-converting enzyme (ACE) is a membrane-anchored ectoprotein that is proteolytically cleaved, yielding an enzymatically active soluble ACE. Two mouse monoclonal antibodies, MAbs 1B3 and 5C8, were generated to the C-terminal part of human soluble ACE. MAb 1B3 recognized the catalytically active ACE, as revealed by ELISA and precipitation assays, whereas Western blotting and immunohistochemisty on paraffin- embedded sections using MAb 5C8 detected denatured ACE. MAb 1B3 showed extensive cross-reactivity, recognizing 15 species out of the 16 tested. The binding of this MAb to ACE was greatly affected by conformational changes induced by adsorption on plastic, formalin fixation, and underglycosylation. Furthermore, MAb 1B3 binding to the mutated ACE (Pro1199Leu substitution in the juxtamembrane region, leading to a fivefold increase in serum ACE level) was markedly decreased. MAb 5C8 detected all the known expression sites of full-size ACE using formalin-fixed and paraffin-embedded human tissues. The sequential epitope for MAb 5C8 is formed by the last 11 amino acid residues of soluble ACE (Pro1193–Arg1203), whereas the conformational epitope for 1B3 is formed by a motif within these 11 amino acid residues and, in addition, by at least one stretch that includes Ala837–His839 located distal to the sequential epitope. Our findings demonstrated that MAbs 1B3 and 5C8 are very useful for the study of ACE shedding, for identification of mutations in stalk regions, and for studying alternatively spliced variants of ACE. In addition, binding of MAb 1B3 is a sensitive determinant of the integrity of soluble ACE. DA - 2005 DB - OpenUCT DP - University of Cape Town J1 - Hybridoma LK - https://open.uct.ac.za PB - University of Cape Town PY - 2005 SM - 1554-0014 T1 - Monoclonal antibodies 1B3 and 5C8 as probes for monitoring the integrity of the C-terminal end of soluble angiotensin-converting enzyme TI - Monoclonal antibodies 1B3 and 5C8 as probes for monitoring the integrity of the C-terminal end of soluble angiotensin-converting enzyme UR - http://hdl.handle.net/11427/21633 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/21633
dc.identifier.vancouvercitationBalyasnikova IV, Sun ZL, Berestetskaya YV, Chubb AJ, Albrecht RF, Sturrock ED, et al. Monoclonal antibodies 1B3 and 5C8 as probes for monitoring the integrity of the C-terminal end of soluble angiotensin-converting enzyme. Hybridoma. 2005; http://hdl.handle.net/11427/21633.en_ZA
dc.languageengen_ZA
dc.publisherMary Ann Lieberten_ZA
dc.publisher.institutionUniversity of Cape Town
dc.sourceHybridomaen_ZA
dc.source.urihttp://www.liebertpub.com/overview/monoclonal-antibodies-brin-immunodiagnosis-brand-immunotherapy/20/
dc.titleMonoclonal antibodies 1B3 and 5C8 as probes for monitoring the integrity of the C-terminal end of soluble angiotensin-converting enzymeen_ZA
dc.typeJournal Articleen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceArticleen_ZA
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