Monoclonal anti-β1-adrenergic receptor antibodies activate G protein signaling in the absence of β-arrestin recruitment
Thermostabilized G protein-coupled receptors used as antigens for in vivo immunization have resulted in the generation of functional agonistic anti-β1-adrenergic (β1AR) receptor monoclonal antibodies (mAbs). The focus of this study was to examine the pharmacology of these antibodies to evaluate thei...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
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Taylor & Francis
2013
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| Online Access: | https://eprints.nottingham.ac.uk/35274/ |
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| author | Hutchings, Catherine J. Cseke, Gabriella Osborne, Greg Woolard, Jeanette Zhukov, Andrei Koglin, Markus Jazayeri, Ali Pandya-Pathak, Jahnavi Langmead, Christopher J. Hill, Stephen J. Weir, Malcolm Marshall, Fiona H. |
| author_facet | Hutchings, Catherine J. Cseke, Gabriella Osborne, Greg Woolard, Jeanette Zhukov, Andrei Koglin, Markus Jazayeri, Ali Pandya-Pathak, Jahnavi Langmead, Christopher J. Hill, Stephen J. Weir, Malcolm Marshall, Fiona H. |
| author_sort | Hutchings, Catherine J. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Thermostabilized G protein-coupled receptors used as antigens for in vivo immunization have resulted in the generation of functional agonistic anti-β1-adrenergic (β1AR) receptor monoclonal antibodies (mAbs). The focus of this study was to examine the pharmacology of these antibodies to evaluate their mechanistic activity at β1AR. Immunization with the β1AR stabilized receptor yielded five stable hybridoma clones, four of which expressed functional IgG, as determined in cell-based assays used to evaluate cAMP stimulation. The antibodies bind diverse epitopes associated with low nanomolar agonist activity at β1AR, and they appeared to show some degree of biased signaling as they were inactive in an assay measuring signaling through β-arrestin. In vitro characterization also verified different antibody-receptor interactions reflecting the different epitopes on the extracellular surface of β1AR to which the mAbs bind. The anti-β1AR mAbs only demonstrated agonist activity when in dimeric antibody format, but not as the monomeric Fab format, suggesting that agonist activation may be mediated through promoting receptor dimerization. Finally, we have also shown that at least one of these antibodies exhibits in vivo functional activity at a therapeutically-relevant dose producing an increase in heart rate consistent with β1AR agonism. |
| first_indexed | 2025-11-14T19:25:47Z |
| format | Article |
| id | nottingham-35274 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:25:47Z |
| publishDate | 2013 |
| publisher | Taylor & Francis |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-352742020-05-04T16:39:46Z https://eprints.nottingham.ac.uk/35274/ Monoclonal anti-β1-adrenergic receptor antibodies activate G protein signaling in the absence of β-arrestin recruitment Hutchings, Catherine J. Cseke, Gabriella Osborne, Greg Woolard, Jeanette Zhukov, Andrei Koglin, Markus Jazayeri, Ali Pandya-Pathak, Jahnavi Langmead, Christopher J. Hill, Stephen J. Weir, Malcolm Marshall, Fiona H. Thermostabilized G protein-coupled receptors used as antigens for in vivo immunization have resulted in the generation of functional agonistic anti-β1-adrenergic (β1AR) receptor monoclonal antibodies (mAbs). The focus of this study was to examine the pharmacology of these antibodies to evaluate their mechanistic activity at β1AR. Immunization with the β1AR stabilized receptor yielded five stable hybridoma clones, four of which expressed functional IgG, as determined in cell-based assays used to evaluate cAMP stimulation. The antibodies bind diverse epitopes associated with low nanomolar agonist activity at β1AR, and they appeared to show some degree of biased signaling as they were inactive in an assay measuring signaling through β-arrestin. In vitro characterization also verified different antibody-receptor interactions reflecting the different epitopes on the extracellular surface of β1AR to which the mAbs bind. The anti-β1AR mAbs only demonstrated agonist activity when in dimeric antibody format, but not as the monomeric Fab format, suggesting that agonist activation may be mediated through promoting receptor dimerization. Finally, we have also shown that at least one of these antibodies exhibits in vivo functional activity at a therapeutically-relevant dose producing an increase in heart rate consistent with β1AR agonism. Taylor & Francis 2013-11-19 Article PeerReviewed Hutchings, Catherine J., Cseke, Gabriella, Osborne, Greg, Woolard, Jeanette, Zhukov, Andrei, Koglin, Markus, Jazayeri, Ali, Pandya-Pathak, Jahnavi, Langmead, Christopher J., Hill, Stephen J., Weir, Malcolm and Marshall, Fiona H. (2013) Monoclonal anti-β1-adrenergic receptor antibodies activate G protein signaling in the absence of β-arrestin recruitment. mAbs, 6 (1). pp. 246-261. ISSN 1942-0870 stabilized receptor Beta 1 adrenergic receptor GPCR extracellular domain extracellular loop functional antibody isoprenaline propranolol http://www.tandfonline.com/doi/abs/10.4161/mabs.27226 doi:10.4161/mabs.27226 doi:10.4161/mabs.27226 |
| spellingShingle | stabilized receptor Beta 1 adrenergic receptor GPCR extracellular domain extracellular loop functional antibody isoprenaline propranolol Hutchings, Catherine J. Cseke, Gabriella Osborne, Greg Woolard, Jeanette Zhukov, Andrei Koglin, Markus Jazayeri, Ali Pandya-Pathak, Jahnavi Langmead, Christopher J. Hill, Stephen J. Weir, Malcolm Marshall, Fiona H. Monoclonal anti-β1-adrenergic receptor antibodies activate G protein signaling in the absence of β-arrestin recruitment |
| title | Monoclonal anti-β1-adrenergic receptor antibodies activate G protein signaling in the absence of β-arrestin recruitment |
| title_full | Monoclonal anti-β1-adrenergic receptor antibodies activate G protein signaling in the absence of β-arrestin recruitment |
| title_fullStr | Monoclonal anti-β1-adrenergic receptor antibodies activate G protein signaling in the absence of β-arrestin recruitment |
| title_full_unstemmed | Monoclonal anti-β1-adrenergic receptor antibodies activate G protein signaling in the absence of β-arrestin recruitment |
| title_short | Monoclonal anti-β1-adrenergic receptor antibodies activate G protein signaling in the absence of β-arrestin recruitment |
| title_sort | monoclonal anti-β1-adrenergic receptor antibodies activate g protein signaling in the absence of β-arrestin recruitment |
| topic | stabilized receptor Beta 1 adrenergic receptor GPCR extracellular domain extracellular loop functional antibody isoprenaline propranolol |
| url | https://eprints.nottingham.ac.uk/35274/ https://eprints.nottingham.ac.uk/35274/ https://eprints.nottingham.ac.uk/35274/ |