Detection of FGF15 in plasma by stable isotope standards and capture by anti-peptide antibodies and targeted mass spectrometry

Takeshi Katafuchi, Daria Esterházy, Andrew Lemoff, Xunshan Ding, Varun Sondhi, Steven A. Kliewer, Hamid Mirzaei, David J. Mangelsdorf

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Fibroblast growth factor 15 (FGF15) has been proposed as a postprandial hormone that signals from intestine to liver to regulate bile acid and carbohydrate homeostasis. However, detecting FGF15 in blood using conventional techniques has proven difficult. Here, we describe a stable isotope standards and capture by anti-peptide antibodies (SISCAPA) assay that combines immuno-enrichment with selected reaction monitoring (SRM) mass spectrometry to overcome this issue. Using this assay, we show that FGF15 circulates in plasma in an FXR and circadian rhythm-dependent manner at concentrations that activate its receptor. Consistent with the proposed endocrine role for FGF15 in liver, mice lacking hepatocyte expression of the obligate FGF15 co-receptor, β-Klotho, have increased bile acid synthesis and reduced glycogen storage despite having supraphysiological plasma FGF15 concentrations. Collectively, these data demonstrate that FGF15 functions as a hormone and highlight the utility of SISCAPA-SRM as a sensitive assay for detecting low-abundance proteins in plasma.

Original languageEnglish (US)
Pages (from-to)898-904
Number of pages7
JournalCell Metabolism
Volume21
Issue number6
DOIs
StatePublished - Jun 2 2015

Fingerprint

Fibroblast Growth Factors
Isotopes
Anti-Idiotypic Antibodies
Mass Spectrometry
Peptides
Bile Acids and Salts
Hormones
Liver
Circadian Rhythm
Glycogen
Intestines
Blood Proteins
Hepatocytes
Homeostasis
Carbohydrates

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Physiology

Cite this

Detection of FGF15 in plasma by stable isotope standards and capture by anti-peptide antibodies and targeted mass spectrometry. / Katafuchi, Takeshi; Esterházy, Daria; Lemoff, Andrew; Ding, Xunshan; Sondhi, Varun; Kliewer, Steven A.; Mirzaei, Hamid; Mangelsdorf, David J.

In: Cell Metabolism, Vol. 21, No. 6, 02.06.2015, p. 898-904.

Research output: Contribution to journalArticle

@article{aa83282c9699482eaddf451a851e43d0,
title = "Detection of FGF15 in plasma by stable isotope standards and capture by anti-peptide antibodies and targeted mass spectrometry",
abstract = "Fibroblast growth factor 15 (FGF15) has been proposed as a postprandial hormone that signals from intestine to liver to regulate bile acid and carbohydrate homeostasis. However, detecting FGF15 in blood using conventional techniques has proven difficult. Here, we describe a stable isotope standards and capture by anti-peptide antibodies (SISCAPA) assay that combines immuno-enrichment with selected reaction monitoring (SRM) mass spectrometry to overcome this issue. Using this assay, we show that FGF15 circulates in plasma in an FXR and circadian rhythm-dependent manner at concentrations that activate its receptor. Consistent with the proposed endocrine role for FGF15 in liver, mice lacking hepatocyte expression of the obligate FGF15 co-receptor, β-Klotho, have increased bile acid synthesis and reduced glycogen storage despite having supraphysiological plasma FGF15 concentrations. Collectively, these data demonstrate that FGF15 functions as a hormone and highlight the utility of SISCAPA-SRM as a sensitive assay for detecting low-abundance proteins in plasma.",
author = "Takeshi Katafuchi and Daria Esterh{\'a}zy and Andrew Lemoff and Xunshan Ding and Varun Sondhi and Kliewer, {Steven A.} and Hamid Mirzaei and Mangelsdorf, {David J.}",
year = "2015",
month = "6",
day = "2",
doi = "10.1016/j.cmet.2015.05.004",
language = "English (US)",
volume = "21",
pages = "898--904",
journal = "Cell Metabolism",
issn = "1550-4131",
publisher = "Cell Press",
number = "6",

}

TY - JOUR

T1 - Detection of FGF15 in plasma by stable isotope standards and capture by anti-peptide antibodies and targeted mass spectrometry

AU - Katafuchi, Takeshi

AU - Esterházy, Daria

AU - Lemoff, Andrew

AU - Ding, Xunshan

AU - Sondhi, Varun

AU - Kliewer, Steven A.

AU - Mirzaei, Hamid

AU - Mangelsdorf, David J.

PY - 2015/6/2

Y1 - 2015/6/2

N2 - Fibroblast growth factor 15 (FGF15) has been proposed as a postprandial hormone that signals from intestine to liver to regulate bile acid and carbohydrate homeostasis. However, detecting FGF15 in blood using conventional techniques has proven difficult. Here, we describe a stable isotope standards and capture by anti-peptide antibodies (SISCAPA) assay that combines immuno-enrichment with selected reaction monitoring (SRM) mass spectrometry to overcome this issue. Using this assay, we show that FGF15 circulates in plasma in an FXR and circadian rhythm-dependent manner at concentrations that activate its receptor. Consistent with the proposed endocrine role for FGF15 in liver, mice lacking hepatocyte expression of the obligate FGF15 co-receptor, β-Klotho, have increased bile acid synthesis and reduced glycogen storage despite having supraphysiological plasma FGF15 concentrations. Collectively, these data demonstrate that FGF15 functions as a hormone and highlight the utility of SISCAPA-SRM as a sensitive assay for detecting low-abundance proteins in plasma.

AB - Fibroblast growth factor 15 (FGF15) has been proposed as a postprandial hormone that signals from intestine to liver to regulate bile acid and carbohydrate homeostasis. However, detecting FGF15 in blood using conventional techniques has proven difficult. Here, we describe a stable isotope standards and capture by anti-peptide antibodies (SISCAPA) assay that combines immuno-enrichment with selected reaction monitoring (SRM) mass spectrometry to overcome this issue. Using this assay, we show that FGF15 circulates in plasma in an FXR and circadian rhythm-dependent manner at concentrations that activate its receptor. Consistent with the proposed endocrine role for FGF15 in liver, mice lacking hepatocyte expression of the obligate FGF15 co-receptor, β-Klotho, have increased bile acid synthesis and reduced glycogen storage despite having supraphysiological plasma FGF15 concentrations. Collectively, these data demonstrate that FGF15 functions as a hormone and highlight the utility of SISCAPA-SRM as a sensitive assay for detecting low-abundance proteins in plasma.

UR - http://www.scopus.com/inward/record.url?scp=84930584701&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84930584701&partnerID=8YFLogxK

U2 - 10.1016/j.cmet.2015.05.004

DO - 10.1016/j.cmet.2015.05.004

M3 - Article

VL - 21

SP - 898

EP - 904

JO - Cell Metabolism

JF - Cell Metabolism

SN - 1550-4131

IS - 6

ER -