Monoclonal Antibodies That Bind to the Ly6 Domain of GPIHBP1 Abolish the Binding of LPL

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

doi:10.1194/jlr.m072462

Monoclonal Antibodies That Bind to the Ly6 Domain of GPIHBP1 Abolish the Binding of LPL

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

14 Citations (Scopus)

Abstract

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.-Hu, X., M. W. Sleeman, K. Miyashita, M. F. Linton, C. M. Allan, C. He, M. Larsson, Y. Tu, N. P. Sandoval, R. S. Jung, A. Mapar, T. Machida, M. Murakami, K. Nakajima, M. Ploug, L. G. Fong, S. G. Young, and A. P. Beigneux. Monoclonal antibodies that bind to the Ly6 domain of GPIHBP1 abolish the binding of LPL.

Original language	English
Journal	Journal of Lipid Research
Volume	58
Pages (from-to)	208-215
Number of pages	10
ISSN	0022-2275
DOIs	https://doi.org/10.1194/jlr.m072462
Publication status	Published - Jan 2017

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Hu, X., Sleeman, M. W., Miyashita, K., Linton, M. F., Allan, C. M., He, C., Larsson, M., Tu, Y., Sandoval, N. P., Jung, R. S., Mapar, A., Machida, T., Murakami, M., Nakajima, K., Ploug, M., Fong, L. G., Young, S. G., & Beigneux, A. P. (2017). Monoclonal Antibodies That Bind to the Ly6 Domain of GPIHBP1 Abolish the Binding of LPL. Journal of Lipid Research, 58, 208-215. https://doi.org/10.1194/jlr.m072462

Hu, X, Sleeman, MW, Miyashita, K, Linton, MF, Allan, CM, He, C, Larsson, M, Tu, Y, Sandoval, NP, Jung, RS, Mapar, A, Machida, T, Murakami, M, Nakajima, K, Ploug, M, Fong, LG, Young, SG & Beigneux, AP 2017, 'Monoclonal Antibodies That Bind to the Ly6 Domain of GPIHBP1 Abolish the Binding of LPL', Journal of Lipid Research, vol. 58, pp. 208-215. https://doi.org/10.1194/jlr.m072462

@article{036d03cf89924f0c9f50a8df8c6813ec,

title = "Monoclonal Antibodies That Bind to the Ly6 Domain of GPIHBP1 Abolish the Binding of LPL",

abstract = "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.-Hu, X., M. W. Sleeman, K. Miyashita, M. F. Linton, C. M. Allan, C. He, M. Larsson, Y. Tu, N. P. Sandoval, R. S. Jung, A. Mapar, T. Machida, M. Murakami, K. Nakajima, M. Ploug, L. G. Fong, S. G. Young, and A. P. Beigneux. Monoclonal antibodies that bind to the Ly6 domain of GPIHBP1 abolish the binding of LPL.",

author = "Xuchen Hu and Sleeman, {Mark W} and Kazuya Miyashita and Linton, {MacRae F} and Allan, {Christopher M} and Cuiwen He and Mikael Larsson and Yiping Tu and Sandoval, {Norma P} and Jung, {Rachel S} and Alaleh Mapar and Tetsuo Machida and Masami Murakami and Katsuyuki Nakajima and Michael Ploug and Fong, {Loren G} and Young, {Stephen G} and Beigneux, {Anne P}",

note = "Copyright {\textcopyright} 2016, The American Society for Biochemistry and Molecular Biology.",

year = "2017",

month = jan,

doi = "10.1194/jlr.m072462",

language = "English",

volume = "58",

pages = "208--215",

journal = "Journal of Lipid Research",

issn = "0022-2275",

publisher = "American Society for Biochemistry and Molecular Biology, Inc.",

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TY - JOUR

T1 - Monoclonal Antibodies That Bind to the Ly6 Domain of GPIHBP1 Abolish the Binding of LPL

AU - Hu, Xuchen

AU - Sleeman, Mark W

AU - Miyashita, Kazuya

AU - Linton, MacRae F

AU - Allan, Christopher M

AU - He, Cuiwen

AU - Larsson, Mikael

AU - Tu, Yiping

AU - Sandoval, Norma P

AU - Jung, Rachel S

AU - Mapar, Alaleh

AU - Machida, Tetsuo

AU - Murakami, Masami

AU - Nakajima, Katsuyuki

AU - Ploug, Michael

AU - Fong, Loren G

AU - Young, Stephen G

AU - Beigneux, Anne P

PY - 2017/1

Y1 - 2017/1

N2 - 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.-Hu, X., M. W. Sleeman, K. Miyashita, M. F. Linton, C. M. Allan, C. He, M. Larsson, Y. Tu, N. P. Sandoval, R. S. Jung, A. Mapar, T. Machida, M. Murakami, K. Nakajima, M. Ploug, L. G. Fong, S. G. Young, and A. P. Beigneux. Monoclonal antibodies that bind to the Ly6 domain of GPIHBP1 abolish the binding of LPL.

AB - 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.-Hu, X., M. W. Sleeman, K. Miyashita, M. F. Linton, C. M. Allan, C. He, M. Larsson, Y. Tu, N. P. Sandoval, R. S. Jung, A. Mapar, T. Machida, M. Murakami, K. Nakajima, M. Ploug, L. G. Fong, S. G. Young, and A. P. Beigneux. Monoclonal antibodies that bind to the Ly6 domain of GPIHBP1 abolish the binding of LPL.

U2 - 10.1194/jlr.m072462

DO - 10.1194/jlr.m072462

M3 - Journal article

C2 - 27875259

SN - 0022-2275

VL - 58

SP - 208

EP - 215

JO - Journal of Lipid Research

JF - Journal of Lipid Research

ER -

Monoclonal Antibodies That Bind to the Ly6 Domain of GPIHBP1 Abolish the Binding of LPL

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