Development and characterisation of a novel glucagon like peptide-1 receptor antibody
Aims/hypothesis Glucagon like peptide-1 (GLP-1) enhances glucose-dependent insulin secretion by binding to GLP-1 receptors (GLP1Rs) on pancreatic beta cells. GLP-1 mimetics are used in the clinic for the treatment of type 2 diabetes, but despite their therapeutic success, several clinical effects of...
Gespeichert in:
| Veröffentlicht in: | Diabetologia 2018-03, Vol.61 (3), p.711-721 |
|---|---|
| Hauptverfasser: | , , , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 721 |
|---|---|
| container_issue | 3 |
| container_start_page | 711 |
| container_title | Diabetologia |
| container_volume | 61 |
| creator | Biggs, Emma K. Liang, Lihuan Naylor, Jacqueline Madalli, Shimona Collier, Rachel Coghlan, Matthew P. Baker, David J. Hornigold, David C. Ravn, Peter Reimann, Frank Gribble, Fiona M. |
| description | Aims/hypothesis
Glucagon like peptide-1 (GLP-1) enhances glucose-dependent insulin secretion by binding to GLP-1 receptors (GLP1Rs) on pancreatic beta cells. GLP-1 mimetics are used in the clinic for the treatment of type 2 diabetes, but despite their therapeutic success, several clinical effects of GLP-1 remain unexplained at a mechanistic level, particularly in extrapancreatic tissues. The aim of this study was to generate and characterise a monoclonal antagonistic antibody for the GLP1R for use in vivo.
Methods
A naive phage display selection strategy was used to isolate single-chain variable fragments (ScFvs) that bound to GLP1R. The ScFv with the highest affinity, Glp1R0017, was converted into a human IgG1 and characterised further. In vitro antagonistic activity was assessed in a number of assays: a cAMP-based homogenous time-resolved fluorescence assay in GLP1R-overexpressing cell lines, a live cell cAMP imaging assay and an insulin secretion assay in INS-1 832/3 cells. Glp1R0017 was further tested in immunostaining of mouse pancreas, and the ability of Glp1R0017 to block GLP1R in vivo was assessed by both IPGTT and OGTT in C57/Bl6 mice.
Results
Antibodies to GLP1R were selected from naive antibody phage display libraries. The monoclonal antibody Glp1R0017 antagonised mouse, human, rat, cynomolgus monkey and dog GLP1R. This antagonistic activity was specific to GLP1R; no antagonistic activity was found in cells overexpressing the glucose-dependent insulinotropic peptide receptor (GIPR), glucagon like peptide-2 receptor or glucagon receptor. GLP-1-stimulated cAMP and insulin secretion was attenuated in INS-1 832/3 cells by Glp1R0017 incubation. Immunostaining of mouse pancreas tissue with Glp1R0017 showed specific staining in the islets of Langerhans, which was absent in
Glp1r
knockout tissue. In vivo, Glp1R0017 reversed the glucose-lowering effect of liraglutide during IPGTTs, and reduced glucose tolerance by blocking endogenous GLP-1 action in OGTTs.
Conclusions/interpretation
Glp1R0017 is a monoclonal antagonistic antibody to the GLP1R that binds to GLP1R on pancreatic beta cells and blocks the actions of GLP-1 in vivo. This antibody holds the potential to be used in investigating the physiological importance of GLP1R signalling in extrapancreatic tissues where cellular targets and signalling pathways activated by GLP-1 are poorly understood. |
| doi_str_mv | 10.1007/s00125-017-4491-0 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5890879</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1962425851</sourcerecordid><originalsourceid>FETCH-LOGICAL-c470t-afa4a93d9fb4bf80a88bf9dd73211f83483f4c92c11f975486f56759fda6466b3</originalsourceid><addsrcrecordid>eNp1kUtLJDEUhYM4TPc48wPcSIEbN5m5SSVVyUYY1HlAgxuF2YVUKulOW10pkyqh_71puhUdcJXH-e5JDgehUwLfCUD9IwEQyjGQGjMmCYYjNCespBgYFcdovpMxEdW_GfqS0hoASs6qz2hGJSGSMj5H99f2yXZh2Nh-LHTfFmalozajjT7p0Ye-CK7QRR8yVSy7yehlvuv8gy0GO4y-tZgU0Zq8DzEbjL4J7fYr-uR0l-y3w3qC7n_d3F39wYvb33-vfi6wYTWMWDvNtCxb6RrWOAFaiMbJtq1LSogTJROlY0ZSk0-y5kxUjlc1l67VFauqpjxBl3vfYWo2tjU5RNSdGqLf6LhVQXv1Xun9Si3Dk-JCgqhlNrg4GMTwONk0qo1Pxnad7m2YkiKyooxywUlGz_9D12GKfY6XKSlBUlmLTJE9ZWJIKVr3-hkCalea2pemcmlqV5qCPHP2NsXrxEtLGaB7IGWpX9r45ukPXZ8B-f2jHA</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1999092978</pqid></control><display><type>article</type><title>Development and characterisation of a novel glucagon like peptide-1 receptor antibody</title><source>MEDLINE</source><source>Springer Nature - Complete Springer Journals</source><creator>Biggs, Emma K. ; Liang, Lihuan ; Naylor, Jacqueline ; Madalli, Shimona ; Collier, Rachel ; Coghlan, Matthew P. ; Baker, David J. ; Hornigold, David C. ; Ravn, Peter ; Reimann, Frank ; Gribble, Fiona M.</creator><creatorcontrib>Biggs, Emma K. ; Liang, Lihuan ; Naylor, Jacqueline ; Madalli, Shimona ; Collier, Rachel ; Coghlan, Matthew P. ; Baker, David J. ; Hornigold, David C. ; Ravn, Peter ; Reimann, Frank ; Gribble, Fiona M.</creatorcontrib><description>Aims/hypothesis
Glucagon like peptide-1 (GLP-1) enhances glucose-dependent insulin secretion by binding to GLP-1 receptors (GLP1Rs) on pancreatic beta cells. GLP-1 mimetics are used in the clinic for the treatment of type 2 diabetes, but despite their therapeutic success, several clinical effects of GLP-1 remain unexplained at a mechanistic level, particularly in extrapancreatic tissues. The aim of this study was to generate and characterise a monoclonal antagonistic antibody for the GLP1R for use in vivo.
Methods
A naive phage display selection strategy was used to isolate single-chain variable fragments (ScFvs) that bound to GLP1R. The ScFv with the highest affinity, Glp1R0017, was converted into a human IgG1 and characterised further. In vitro antagonistic activity was assessed in a number of assays: a cAMP-based homogenous time-resolved fluorescence assay in GLP1R-overexpressing cell lines, a live cell cAMP imaging assay and an insulin secretion assay in INS-1 832/3 cells. Glp1R0017 was further tested in immunostaining of mouse pancreas, and the ability of Glp1R0017 to block GLP1R in vivo was assessed by both IPGTT and OGTT in C57/Bl6 mice.
Results
Antibodies to GLP1R were selected from naive antibody phage display libraries. The monoclonal antibody Glp1R0017 antagonised mouse, human, rat, cynomolgus monkey and dog GLP1R. This antagonistic activity was specific to GLP1R; no antagonistic activity was found in cells overexpressing the glucose-dependent insulinotropic peptide receptor (GIPR), glucagon like peptide-2 receptor or glucagon receptor. GLP-1-stimulated cAMP and insulin secretion was attenuated in INS-1 832/3 cells by Glp1R0017 incubation. Immunostaining of mouse pancreas tissue with Glp1R0017 showed specific staining in the islets of Langerhans, which was absent in
Glp1r
knockout tissue. In vivo, Glp1R0017 reversed the glucose-lowering effect of liraglutide during IPGTTs, and reduced glucose tolerance by blocking endogenous GLP-1 action in OGTTs.
Conclusions/interpretation
Glp1R0017 is a monoclonal antagonistic antibody to the GLP1R that binds to GLP1R on pancreatic beta cells and blocks the actions of GLP-1 in vivo. This antibody holds the potential to be used in investigating the physiological importance of GLP1R signalling in extrapancreatic tissues where cellular targets and signalling pathways activated by GLP-1 are poorly understood.</description><identifier>ISSN: 0012-186X</identifier><identifier>EISSN: 1432-0428</identifier><identifier>DOI: 10.1007/s00125-017-4491-0</identifier><identifier>PMID: 29119245</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Animals ; Antibodies - immunology ; Beta cells ; Calcium - metabolism ; Cell Line ; Cell lines ; CHO Cells ; Cricetulus ; Cyclic AMP ; Cyclic AMP - metabolism ; Diabetes mellitus ; Diabetes mellitus (non-insulin dependent) ; Glucagon ; Glucagon-Like Peptide-1 Receptor - antagonists & inhibitors ; Glucagon-Like Peptide-1 Receptor - immunology ; Glucose ; Glucose tolerance ; HEK293 Cells ; Human Physiology ; Humans ; Immunoglobulin G ; Immunoglobulin G - metabolism ; Immunological tolerance ; Incretins - metabolism ; Insulin ; Insulin - metabolism ; Insulin secretion ; Internal Medicine ; Islets of Langerhans ; Medicine ; Medicine & Public Health ; Metabolic Diseases ; Mice ; Monoclonal antibodies ; Pancreas ; Peptide Library ; Peptides ; Phage display ; Signal transduction</subject><ispartof>Diabetologia, 2018-03, Vol.61 (3), p.711-721</ispartof><rights>The Author(s) 2017</rights><rights>Diabetologia is a copyright of Springer, (2017). All Rights Reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c470t-afa4a93d9fb4bf80a88bf9dd73211f83483f4c92c11f975486f56759fda6466b3</citedby><cites>FETCH-LOGICAL-c470t-afa4a93d9fb4bf80a88bf9dd73211f83483f4c92c11f975486f56759fda6466b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00125-017-4491-0$$EPDF$$P50$$Gspringer$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00125-017-4491-0$$EHTML$$P50$$Gspringer$$Hfree_for_read</linktohtml><link.rule.ids>230,314,776,780,881,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29119245$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Biggs, Emma K.</creatorcontrib><creatorcontrib>Liang, Lihuan</creatorcontrib><creatorcontrib>Naylor, Jacqueline</creatorcontrib><creatorcontrib>Madalli, Shimona</creatorcontrib><creatorcontrib>Collier, Rachel</creatorcontrib><creatorcontrib>Coghlan, Matthew P.</creatorcontrib><creatorcontrib>Baker, David J.</creatorcontrib><creatorcontrib>Hornigold, David C.</creatorcontrib><creatorcontrib>Ravn, Peter</creatorcontrib><creatorcontrib>Reimann, Frank</creatorcontrib><creatorcontrib>Gribble, Fiona M.</creatorcontrib><title>Development and characterisation of a novel glucagon like peptide-1 receptor antibody</title><title>Diabetologia</title><addtitle>Diabetologia</addtitle><addtitle>Diabetologia</addtitle><description>Aims/hypothesis
Glucagon like peptide-1 (GLP-1) enhances glucose-dependent insulin secretion by binding to GLP-1 receptors (GLP1Rs) on pancreatic beta cells. GLP-1 mimetics are used in the clinic for the treatment of type 2 diabetes, but despite their therapeutic success, several clinical effects of GLP-1 remain unexplained at a mechanistic level, particularly in extrapancreatic tissues. The aim of this study was to generate and characterise a monoclonal antagonistic antibody for the GLP1R for use in vivo.
Methods
A naive phage display selection strategy was used to isolate single-chain variable fragments (ScFvs) that bound to GLP1R. The ScFv with the highest affinity, Glp1R0017, was converted into a human IgG1 and characterised further. In vitro antagonistic activity was assessed in a number of assays: a cAMP-based homogenous time-resolved fluorescence assay in GLP1R-overexpressing cell lines, a live cell cAMP imaging assay and an insulin secretion assay in INS-1 832/3 cells. Glp1R0017 was further tested in immunostaining of mouse pancreas, and the ability of Glp1R0017 to block GLP1R in vivo was assessed by both IPGTT and OGTT in C57/Bl6 mice.
Results
Antibodies to GLP1R were selected from naive antibody phage display libraries. The monoclonal antibody Glp1R0017 antagonised mouse, human, rat, cynomolgus monkey and dog GLP1R. This antagonistic activity was specific to GLP1R; no antagonistic activity was found in cells overexpressing the glucose-dependent insulinotropic peptide receptor (GIPR), glucagon like peptide-2 receptor or glucagon receptor. GLP-1-stimulated cAMP and insulin secretion was attenuated in INS-1 832/3 cells by Glp1R0017 incubation. Immunostaining of mouse pancreas tissue with Glp1R0017 showed specific staining in the islets of Langerhans, which was absent in
Glp1r
knockout tissue. In vivo, Glp1R0017 reversed the glucose-lowering effect of liraglutide during IPGTTs, and reduced glucose tolerance by blocking endogenous GLP-1 action in OGTTs.
Conclusions/interpretation
Glp1R0017 is a monoclonal antagonistic antibody to the GLP1R that binds to GLP1R on pancreatic beta cells and blocks the actions of GLP-1 in vivo. This antibody holds the potential to be used in investigating the physiological importance of GLP1R signalling in extrapancreatic tissues where cellular targets and signalling pathways activated by GLP-1 are poorly understood.</description><subject>Animals</subject><subject>Antibodies - immunology</subject><subject>Beta cells</subject><subject>Calcium - metabolism</subject><subject>Cell Line</subject><subject>Cell lines</subject><subject>CHO Cells</subject><subject>Cricetulus</subject><subject>Cyclic AMP</subject><subject>Cyclic AMP - metabolism</subject><subject>Diabetes mellitus</subject><subject>Diabetes mellitus (non-insulin dependent)</subject><subject>Glucagon</subject><subject>Glucagon-Like Peptide-1 Receptor - antagonists & inhibitors</subject><subject>Glucagon-Like Peptide-1 Receptor - immunology</subject><subject>Glucose</subject><subject>Glucose tolerance</subject><subject>HEK293 Cells</subject><subject>Human Physiology</subject><subject>Humans</subject><subject>Immunoglobulin G</subject><subject>Immunoglobulin G - metabolism</subject><subject>Immunological tolerance</subject><subject>Incretins - metabolism</subject><subject>Insulin</subject><subject>Insulin - metabolism</subject><subject>Insulin secretion</subject><subject>Internal Medicine</subject><subject>Islets of Langerhans</subject><subject>Medicine</subject><subject>Medicine & Public Health</subject><subject>Metabolic Diseases</subject><subject>Mice</subject><subject>Monoclonal antibodies</subject><subject>Pancreas</subject><subject>Peptide Library</subject><subject>Peptides</subject><subject>Phage display</subject><subject>Signal transduction</subject><issn>0012-186X</issn><issn>1432-0428</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNp1kUtLJDEUhYM4TPc48wPcSIEbN5m5SSVVyUYY1HlAgxuF2YVUKulOW10pkyqh_71puhUdcJXH-e5JDgehUwLfCUD9IwEQyjGQGjMmCYYjNCespBgYFcdovpMxEdW_GfqS0hoASs6qz2hGJSGSMj5H99f2yXZh2Nh-LHTfFmalozajjT7p0Ye-CK7QRR8yVSy7yehlvuv8gy0GO4y-tZgU0Zq8DzEbjL4J7fYr-uR0l-y3w3qC7n_d3F39wYvb33-vfi6wYTWMWDvNtCxb6RrWOAFaiMbJtq1LSogTJROlY0ZSk0-y5kxUjlc1l67VFauqpjxBl3vfYWo2tjU5RNSdGqLf6LhVQXv1Xun9Si3Dk-JCgqhlNrg4GMTwONk0qo1Pxnad7m2YkiKyooxywUlGz_9D12GKfY6XKSlBUlmLTJE9ZWJIKVr3-hkCalea2pemcmlqV5qCPHP2NsXrxEtLGaB7IGWpX9r45ukPXZ8B-f2jHA</recordid><startdate>20180301</startdate><enddate>20180301</enddate><creator>Biggs, Emma K.</creator><creator>Liang, Lihuan</creator><creator>Naylor, Jacqueline</creator><creator>Madalli, Shimona</creator><creator>Collier, Rachel</creator><creator>Coghlan, Matthew P.</creator><creator>Baker, David J.</creator><creator>Hornigold, David C.</creator><creator>Ravn, Peter</creator><creator>Reimann, Frank</creator><creator>Gribble, Fiona M.</creator><general>Springer Berlin Heidelberg</general><general>Springer Nature B.V</general><scope>C6C</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7T5</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>H94</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>PHGZM</scope><scope>PHGZT</scope><scope>PJZUB</scope><scope>PKEHL</scope><scope>PPXIY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20180301</creationdate><title>Development and characterisation of a novel glucagon like peptide-1 receptor antibody</title><author>Biggs, Emma K. ; Liang, Lihuan ; Naylor, Jacqueline ; Madalli, Shimona ; Collier, Rachel ; Coghlan, Matthew P. ; Baker, David J. ; Hornigold, David C. ; Ravn, Peter ; Reimann, Frank ; Gribble, Fiona M.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c470t-afa4a93d9fb4bf80a88bf9dd73211f83483f4c92c11f975486f56759fda6466b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Animals</topic><topic>Antibodies - immunology</topic><topic>Beta cells</topic><topic>Calcium - metabolism</topic><topic>Cell Line</topic><topic>Cell lines</topic><topic>CHO Cells</topic><topic>Cricetulus</topic><topic>Cyclic AMP</topic><topic>Cyclic AMP - metabolism</topic><topic>Diabetes mellitus</topic><topic>Diabetes mellitus (non-insulin dependent)</topic><topic>Glucagon</topic><topic>Glucagon-Like Peptide-1 Receptor - antagonists & inhibitors</topic><topic>Glucagon-Like Peptide-1 Receptor - immunology</topic><topic>Glucose</topic><topic>Glucose tolerance</topic><topic>HEK293 Cells</topic><topic>Human Physiology</topic><topic>Humans</topic><topic>Immunoglobulin G</topic><topic>Immunoglobulin G - metabolism</topic><topic>Immunological tolerance</topic><topic>Incretins - metabolism</topic><topic>Insulin</topic><topic>Insulin - metabolism</topic><topic>Insulin secretion</topic><topic>Internal Medicine</topic><topic>Islets of Langerhans</topic><topic>Medicine</topic><topic>Medicine & Public Health</topic><topic>Metabolic Diseases</topic><topic>Mice</topic><topic>Monoclonal antibodies</topic><topic>Pancreas</topic><topic>Peptide Library</topic><topic>Peptides</topic><topic>Phage display</topic><topic>Signal transduction</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Biggs, Emma K.</creatorcontrib><creatorcontrib>Liang, Lihuan</creatorcontrib><creatorcontrib>Naylor, Jacqueline</creatorcontrib><creatorcontrib>Madalli, Shimona</creatorcontrib><creatorcontrib>Collier, Rachel</creatorcontrib><creatorcontrib>Coghlan, Matthew P.</creatorcontrib><creatorcontrib>Baker, David J.</creatorcontrib><creatorcontrib>Hornigold, David C.</creatorcontrib><creatorcontrib>Ravn, Peter</creatorcontrib><creatorcontrib>Reimann, Frank</creatorcontrib><creatorcontrib>Gribble, Fiona M.</creatorcontrib><collection>Springer Nature OA Free Journals</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Immunology Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>ProQuest Central (New)</collection><collection>ProQuest One Academic (New)</collection><collection>ProQuest Health & Medical Research Collection</collection><collection>ProQuest One Academic Middle East (New)</collection><collection>ProQuest One Health & Nursing</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Diabetologia</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Biggs, Emma K.</au><au>Liang, Lihuan</au><au>Naylor, Jacqueline</au><au>Madalli, Shimona</au><au>Collier, Rachel</au><au>Coghlan, Matthew P.</au><au>Baker, David J.</au><au>Hornigold, David C.</au><au>Ravn, Peter</au><au>Reimann, Frank</au><au>Gribble, Fiona M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development and characterisation of a novel glucagon like peptide-1 receptor antibody</atitle><jtitle>Diabetologia</jtitle><stitle>Diabetologia</stitle><addtitle>Diabetologia</addtitle><date>2018-03-01</date><risdate>2018</risdate><volume>61</volume><issue>3</issue><spage>711</spage><epage>721</epage><pages>711-721</pages><issn>0012-186X</issn><eissn>1432-0428</eissn><abstract>Aims/hypothesis
Glucagon like peptide-1 (GLP-1) enhances glucose-dependent insulin secretion by binding to GLP-1 receptors (GLP1Rs) on pancreatic beta cells. GLP-1 mimetics are used in the clinic for the treatment of type 2 diabetes, but despite their therapeutic success, several clinical effects of GLP-1 remain unexplained at a mechanistic level, particularly in extrapancreatic tissues. The aim of this study was to generate and characterise a monoclonal antagonistic antibody for the GLP1R for use in vivo.
Methods
A naive phage display selection strategy was used to isolate single-chain variable fragments (ScFvs) that bound to GLP1R. The ScFv with the highest affinity, Glp1R0017, was converted into a human IgG1 and characterised further. In vitro antagonistic activity was assessed in a number of assays: a cAMP-based homogenous time-resolved fluorescence assay in GLP1R-overexpressing cell lines, a live cell cAMP imaging assay and an insulin secretion assay in INS-1 832/3 cells. Glp1R0017 was further tested in immunostaining of mouse pancreas, and the ability of Glp1R0017 to block GLP1R in vivo was assessed by both IPGTT and OGTT in C57/Bl6 mice.
Results
Antibodies to GLP1R were selected from naive antibody phage display libraries. The monoclonal antibody Glp1R0017 antagonised mouse, human, rat, cynomolgus monkey and dog GLP1R. This antagonistic activity was specific to GLP1R; no antagonistic activity was found in cells overexpressing the glucose-dependent insulinotropic peptide receptor (GIPR), glucagon like peptide-2 receptor or glucagon receptor. GLP-1-stimulated cAMP and insulin secretion was attenuated in INS-1 832/3 cells by Glp1R0017 incubation. Immunostaining of mouse pancreas tissue with Glp1R0017 showed specific staining in the islets of Langerhans, which was absent in
Glp1r
knockout tissue. In vivo, Glp1R0017 reversed the glucose-lowering effect of liraglutide during IPGTTs, and reduced glucose tolerance by blocking endogenous GLP-1 action in OGTTs.
Conclusions/interpretation
Glp1R0017 is a monoclonal antagonistic antibody to the GLP1R that binds to GLP1R on pancreatic beta cells and blocks the actions of GLP-1 in vivo. This antibody holds the potential to be used in investigating the physiological importance of GLP1R signalling in extrapancreatic tissues where cellular targets and signalling pathways activated by GLP-1 are poorly understood.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>29119245</pmid><doi>10.1007/s00125-017-4491-0</doi><tpages>11</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0012-186X |
| ispartof | Diabetologia, 2018-03, Vol.61 (3), p.711-721 |
| issn | 0012-186X 1432-0428 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5890879 |
| source | MEDLINE; Springer Nature - Complete Springer Journals |
| subjects | Animals Antibodies - immunology Beta cells Calcium - metabolism Cell Line Cell lines CHO Cells Cricetulus Cyclic AMP Cyclic AMP - metabolism Diabetes mellitus Diabetes mellitus (non-insulin dependent) Glucagon Glucagon-Like Peptide-1 Receptor - antagonists & inhibitors Glucagon-Like Peptide-1 Receptor - immunology Glucose Glucose tolerance HEK293 Cells Human Physiology Humans Immunoglobulin G Immunoglobulin G - metabolism Immunological tolerance Incretins - metabolism Insulin Insulin - metabolism Insulin secretion Internal Medicine Islets of Langerhans Medicine Medicine & Public Health Metabolic Diseases Mice Monoclonal antibodies Pancreas Peptide Library Peptides Phage display Signal transduction |
| title | Development and characterisation of a novel glucagon like peptide-1 receptor antibody |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-18T21%3A00%3A21IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Development%20and%20characterisation%20of%20a%20novel%20glucagon%20like%20peptide-1%20receptor%20antibody&rft.jtitle=Diabetologia&rft.au=Biggs,%20Emma%20K.&rft.date=2018-03-01&rft.volume=61&rft.issue=3&rft.spage=711&rft.epage=721&rft.pages=711-721&rft.issn=0012-186X&rft.eissn=1432-0428&rft_id=info:doi/10.1007/s00125-017-4491-0&rft_dat=%3Cproquest_pubme%3E1962425851%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1999092978&rft_id=info:pmid/29119245&rfr_iscdi=true |