Monoclonal antibodies that bind to the Ly6 domain of GPIHBP1 abolish the binding of LPL
GPIHBP1, an endothelial cell protein, binds LPL in the interstitial spaces and shuttles it to its site of action inside blood vessels. For years, studies of human GPIHBP1 have been hampered by an absence of useful antibodies. We reasoned that monoclonal antibodies (mAbs) against human GPIHBP1 would...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2017-01-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520314486 |
| _version_ | 1818726425357713408 |
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| author | Xuchen Hu Mark W. Sleeman Kazuya Miyashita MacRae F. Linton Christopher M. Allan Cuiwen He Mikael Larsson Yiping Tu Norma P. Sandoval Rachel S. Jung Alaleh Mapar Tetsuo Machida Masami Murakami Katsuyuki Nakajima Michael Ploug Loren G. Fong Stephen G. Young Anne P. Beigneux |
| author_facet | Xuchen Hu Mark W. Sleeman Kazuya Miyashita MacRae F. Linton Christopher M. Allan Cuiwen He Mikael Larsson Yiping Tu Norma P. Sandoval Rachel S. Jung Alaleh Mapar Tetsuo Machida Masami Murakami Katsuyuki Nakajima Michael Ploug Loren G. Fong Stephen G. Young Anne P. Beigneux |
| author_sort | Xuchen Hu |
| collection | DOAJ |
| description | GPIHBP1, an endothelial cell protein, binds LPL in the interstitial spaces and shuttles it to its site of action inside blood vessels. For years, studies of human GPIHBP1 have been hampered by an absence of useful antibodies. We reasoned that monoclonal antibodies (mAbs) against human GPIHBP1 would be useful for 1) defining the functional relevance of GPIHBP1's Ly6 and acidic domains to the binding of LPL; 2) ascertaining whether human GPIHBP1 is expressed exclusively in capillary endothelial cells; and 3) testing whether GPIHBP1 is detectable in human plasma. Here, we report the development of a panel of human GPIHBP1-specific mAbs. Two mAbs against GPIHBP1's Ly6 domain, RE3 and RG3, abolished LPL binding, whereas an antibody against the acidic domain, RF4, did not. Also, mAbs RE3 and RG3 bound with reduced affinity to a mutant GPIHBP1 containing an Ly6 domain mutation (W109S) that abolishes LPL binding. Immunohistochemistry studies with the GPIHBP1 mAbs revealed that human GPIHBP1 is expressed only in capillary endothelial cells. Finally, we created an ELISA that detects GPIHBP1 in human plasma. That ELISA should make it possible for clinical lipidologists to determine whether plasma GPIHBP1 levels are a useful biomarker of metabolic or vascular disease |
| first_indexed | 2024-12-17T21:58:00Z |
| format | Article |
| id | doaj.art-2cd466d9dd7b46b5858c18eb7905f80f |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-17T21:58:00Z |
| publishDate | 2017-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-2cd466d9dd7b46b5858c18eb7905f80f2022-12-21T21:31:04ZengElsevierJournal of Lipid Research0022-22752017-01-01581208215Monoclonal antibodies that bind to the Ly6 domain of GPIHBP1 abolish the binding of LPLXuchen Hu0Mark W. Sleeman1Kazuya Miyashita2MacRae F. Linton3Christopher M. Allan4Cuiwen He5Mikael Larsson6Yiping Tu7Norma P. Sandoval8Rachel S. Jung9Alaleh Mapar10Tetsuo Machida11Masami Murakami12Katsuyuki Nakajima13Michael Ploug14Loren G. Fong15Stephen G. Young16Anne P. Beigneux17Departments of Medicine University of California Los Angeles, Los Angeles, CAMonash Biomedicine Discovery Institute and Antibody Technologies Facility, Monash University, Victoria, AustraliaDepartment of Clinical Laboratory Medicine, Graduate School of Medicine, Gunma University, Maebashi, JapanDepartments of Medicine and Pharmacology, Vanderbilt University Medical Center, Nashville, TNDepartments of Medicine University of California Los Angeles, Los Angeles, CADepartments of Medicine University of California Los Angeles, Los Angeles, CADepartments of Medicine University of California Los Angeles, Los Angeles, CADepartments of Medicine University of California Los Angeles, Los Angeles, CADepartments of Medicine University of California Los Angeles, Los Angeles, CADepartments of Medicine University of California Los Angeles, Los Angeles, CADepartments of Medicine University of California Los Angeles, Los Angeles, CADepartment of Clinical Laboratory Medicine, Graduate School of Medicine, Gunma University, Maebashi, JapanDepartment of Clinical Laboratory Medicine, Graduate School of Medicine, Gunma University, Maebashi, JapanDepartment of Clinical Laboratory Medicine, Graduate School of Medicine, Gunma University, Maebashi, JapanFinsen Laboratory, Rigshospitalet, Copenhagen, Denmark; Biotech Research and Innovation Centre, University of Copenhagen, Copenhagen, DenmarkDepartments of Medicine University of California Los Angeles, Los Angeles, CA; To whom correspondence should be addressed.Departments of Medicine University of California Los Angeles, Los Angeles, CA; Human Genetics, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA; To whom correspondence should be addressed.Departments of Medicine University of California Los Angeles, Los Angeles, CA; To whom correspondence should be addressed.GPIHBP1, an endothelial cell protein, binds LPL in the interstitial spaces and shuttles it to its site of action inside blood vessels. For years, studies of human GPIHBP1 have been hampered by an absence of useful antibodies. We reasoned that monoclonal antibodies (mAbs) against human GPIHBP1 would be useful for 1) defining the functional relevance of GPIHBP1's Ly6 and acidic domains to the binding of LPL; 2) ascertaining whether human GPIHBP1 is expressed exclusively in capillary endothelial cells; and 3) testing whether GPIHBP1 is detectable in human plasma. Here, we report the development of a panel of human GPIHBP1-specific mAbs. Two mAbs against GPIHBP1's Ly6 domain, RE3 and RG3, abolished LPL binding, whereas an antibody against the acidic domain, RF4, did not. Also, mAbs RE3 and RG3 bound with reduced affinity to a mutant GPIHBP1 containing an Ly6 domain mutation (W109S) that abolishes LPL binding. Immunohistochemistry studies with the GPIHBP1 mAbs revealed that human GPIHBP1 is expressed only in capillary endothelial cells. Finally, we created an ELISA that detects GPIHBP1 in human plasma. That ELISA should make it possible for clinical lipidologists to determine whether plasma GPIHBP1 levels are a useful biomarker of metabolic or vascular diseasehttp://www.sciencedirect.com/science/article/pii/S0022227520314486chylomicronsdyslipidemiasendothelial cellslipoprotein lipasetriglycerides |
| spellingShingle | Xuchen Hu Mark W. Sleeman Kazuya Miyashita MacRae F. Linton Christopher M. Allan Cuiwen He Mikael Larsson Yiping Tu Norma P. Sandoval Rachel S. Jung Alaleh Mapar Tetsuo Machida Masami Murakami Katsuyuki Nakajima Michael Ploug Loren G. Fong Stephen G. Young Anne P. Beigneux Monoclonal antibodies that bind to the Ly6 domain of GPIHBP1 abolish the binding of LPL Journal of Lipid Research chylomicrons dyslipidemias endothelial cells lipoprotein lipase triglycerides |
| title | Monoclonal antibodies that bind to the Ly6 domain of GPIHBP1 abolish the binding of LPL |
| title_full | Monoclonal antibodies that bind to the Ly6 domain of GPIHBP1 abolish the binding of LPL |
| title_fullStr | Monoclonal antibodies that bind to the Ly6 domain of GPIHBP1 abolish the binding of LPL |
| title_full_unstemmed | Monoclonal antibodies that bind to the Ly6 domain of GPIHBP1 abolish the binding of LPL |
| title_short | Monoclonal antibodies that bind to the Ly6 domain of GPIHBP1 abolish the binding of LPL |
| title_sort | monoclonal antibodies that bind to the ly6 domain of gpihbp1 abolish the binding of lpl |
| topic | chylomicrons dyslipidemias endothelial cells lipoprotein lipase triglycerides |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520314486 |
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