A novel, multiplexed targeted mass spectrometry assay for quantification of complement factor H (CFH) variants and CFH-related proteins 1–5 in human plasma

Pingbo Zhang, Min Zhu, Minghui Geng-Spyropoulos, Michelle Shardell, Marta Gonzalez-Freire, Vilmundur Gudnason, Gudny Eiriksdottir, Debra Schaumberg, Jennifer E. Van Eyk, Luigi Ferrucci, Richard David Semba

Research output: Contribution to journalArticle

Abstract

Age-related macular degeneration (AMD) is a leading cause of visual loss among older adults. Two variants in the complement factor H (CFH) gene, Y402H and I62V, are strongly associated with risk of AMD. CFH is encoded in regulator of complement activation gene cluster in chromosome 1q32, which includes complement factor related (CFHR) proteins, CFHR1 to CFHR5, with high amino acid sequence homology to CFH. Our goal was to build a SRM assay to measure plasma concentrations of CFH variants Y402, H402, I62, and V62, and CFHR1-5. The final assay consisted of 24 peptides and 72 interference-free SRM transition ion pairs. Most peptides showed good linearity over 0.3–200 fmol/μL concentration range. Plasma concentrations of CFH variants and CFHR1-5 were measured using the SRM assay in 344 adults. Plasma CFH concentrations (mean, SE in μg/mL) by inferred genotype were: YY402, II62 (170.1, 31.4), YY402, VV62 (188.8, 38.5), HH402, VV62 (144.0, 37.0), HY402, VV62 (164.2, 42.3), YY402, IV62 (194.8, 36.8), HY402, IV62 (181.3, 44.7). Mean (SE) plasma concentrations of CFHR1-5 were 1.63 (0.04), 3.64 (1.20), 0.020 (0.001), 2.42 (0.18), and 5.49 (1.55) μg/mL, respectively. This SRM assay should facilitate the study of the role of systemic complement and risk of AMD.

Original languageEnglish (US)
Article number1600237
JournalProteomics
Volume17
Issue number6
DOIs
StatePublished - Mar 1 2017

Fingerprint

Plasma (human)
Complement Factor H
Mass spectrometry
Assays
Mass Spectrometry
Macular Degeneration
Plasmas
Proteins
Genes
Amino Acid Sequence Homology
Peptides
Complement Activation
Chromosomes
Multigene Family
Chemical activation
Genotype
Ions
Amino Acids

Keywords

  • Age-related macular degeneration
  • Biomedicine
  • Complement factor H
  • Complement factor H-related proteins
  • Mass spectrometry
  • Selected reaction monitoring

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

A novel, multiplexed targeted mass spectrometry assay for quantification of complement factor H (CFH) variants and CFH-related proteins 1–5 in human plasma. / Zhang, Pingbo; Zhu, Min; Geng-Spyropoulos, Minghui; Shardell, Michelle; Gonzalez-Freire, Marta; Gudnason, Vilmundur; Eiriksdottir, Gudny; Schaumberg, Debra; Van Eyk, Jennifer E.; Ferrucci, Luigi; Semba, Richard David.

In: Proteomics, Vol. 17, No. 6, 1600237, 01.03.2017.

Research output: Contribution to journalArticle

Zhang, P, Zhu, M, Geng-Spyropoulos, M, Shardell, M, Gonzalez-Freire, M, Gudnason, V, Eiriksdottir, G, Schaumberg, D, Van Eyk, JE, Ferrucci, L & Semba, RD 2017, 'A novel, multiplexed targeted mass spectrometry assay for quantification of complement factor H (CFH) variants and CFH-related proteins 1–5 in human plasma', Proteomics, vol. 17, no. 6, 1600237. https://doi.org/10.1002/pmic.201600237
Zhang, Pingbo ; Zhu, Min ; Geng-Spyropoulos, Minghui ; Shardell, Michelle ; Gonzalez-Freire, Marta ; Gudnason, Vilmundur ; Eiriksdottir, Gudny ; Schaumberg, Debra ; Van Eyk, Jennifer E. ; Ferrucci, Luigi ; Semba, Richard David. / A novel, multiplexed targeted mass spectrometry assay for quantification of complement factor H (CFH) variants and CFH-related proteins 1–5 in human plasma. In: Proteomics. 2017 ; Vol. 17, No. 6.
@article{e2e41ec9e9ce4ff9ab67b82607868359,
title = "A novel, multiplexed targeted mass spectrometry assay for quantification of complement factor H (CFH) variants and CFH-related proteins 1–5 in human plasma",
abstract = "Age-related macular degeneration (AMD) is a leading cause of visual loss among older adults. Two variants in the complement factor H (CFH) gene, Y402H and I62V, are strongly associated with risk of AMD. CFH is encoded in regulator of complement activation gene cluster in chromosome 1q32, which includes complement factor related (CFHR) proteins, CFHR1 to CFHR5, with high amino acid sequence homology to CFH. Our goal was to build a SRM assay to measure plasma concentrations of CFH variants Y402, H402, I62, and V62, and CFHR1-5. The final assay consisted of 24 peptides and 72 interference-free SRM transition ion pairs. Most peptides showed good linearity over 0.3–200 fmol/μL concentration range. Plasma concentrations of CFH variants and CFHR1-5 were measured using the SRM assay in 344 adults. Plasma CFH concentrations (mean, SE in μg/mL) by inferred genotype were: YY402, II62 (170.1, 31.4), YY402, VV62 (188.8, 38.5), HH402, VV62 (144.0, 37.0), HY402, VV62 (164.2, 42.3), YY402, IV62 (194.8, 36.8), HY402, IV62 (181.3, 44.7). Mean (SE) plasma concentrations of CFHR1-5 were 1.63 (0.04), 3.64 (1.20), 0.020 (0.001), 2.42 (0.18), and 5.49 (1.55) μg/mL, respectively. This SRM assay should facilitate the study of the role of systemic complement and risk of AMD.",
keywords = "Age-related macular degeneration, Biomedicine, Complement factor H, Complement factor H-related proteins, Mass spectrometry, Selected reaction monitoring",
author = "Pingbo Zhang and Min Zhu and Minghui Geng-Spyropoulos and Michelle Shardell and Marta Gonzalez-Freire and Vilmundur Gudnason and Gudny Eiriksdottir and Debra Schaumberg and {Van Eyk}, {Jennifer E.} and Luigi Ferrucci and Semba, {Richard David}",
year = "2017",
month = "3",
day = "1",
doi = "10.1002/pmic.201600237",
language = "English (US)",
volume = "17",
journal = "Proteomics",
issn = "1615-9853",
publisher = "Wiley-VCH Verlag",
number = "6",

}

TY - JOUR

T1 - A novel, multiplexed targeted mass spectrometry assay for quantification of complement factor H (CFH) variants and CFH-related proteins 1–5 in human plasma

AU - Zhang, Pingbo

AU - Zhu, Min

AU - Geng-Spyropoulos, Minghui

AU - Shardell, Michelle

AU - Gonzalez-Freire, Marta

AU - Gudnason, Vilmundur

AU - Eiriksdottir, Gudny

AU - Schaumberg, Debra

AU - Van Eyk, Jennifer E.

AU - Ferrucci, Luigi

AU - Semba, Richard David

PY - 2017/3/1

Y1 - 2017/3/1

N2 - Age-related macular degeneration (AMD) is a leading cause of visual loss among older adults. Two variants in the complement factor H (CFH) gene, Y402H and I62V, are strongly associated with risk of AMD. CFH is encoded in regulator of complement activation gene cluster in chromosome 1q32, which includes complement factor related (CFHR) proteins, CFHR1 to CFHR5, with high amino acid sequence homology to CFH. Our goal was to build a SRM assay to measure plasma concentrations of CFH variants Y402, H402, I62, and V62, and CFHR1-5. The final assay consisted of 24 peptides and 72 interference-free SRM transition ion pairs. Most peptides showed good linearity over 0.3–200 fmol/μL concentration range. Plasma concentrations of CFH variants and CFHR1-5 were measured using the SRM assay in 344 adults. Plasma CFH concentrations (mean, SE in μg/mL) by inferred genotype were: YY402, II62 (170.1, 31.4), YY402, VV62 (188.8, 38.5), HH402, VV62 (144.0, 37.0), HY402, VV62 (164.2, 42.3), YY402, IV62 (194.8, 36.8), HY402, IV62 (181.3, 44.7). Mean (SE) plasma concentrations of CFHR1-5 were 1.63 (0.04), 3.64 (1.20), 0.020 (0.001), 2.42 (0.18), and 5.49 (1.55) μg/mL, respectively. This SRM assay should facilitate the study of the role of systemic complement and risk of AMD.

AB - Age-related macular degeneration (AMD) is a leading cause of visual loss among older adults. Two variants in the complement factor H (CFH) gene, Y402H and I62V, are strongly associated with risk of AMD. CFH is encoded in regulator of complement activation gene cluster in chromosome 1q32, which includes complement factor related (CFHR) proteins, CFHR1 to CFHR5, with high amino acid sequence homology to CFH. Our goal was to build a SRM assay to measure plasma concentrations of CFH variants Y402, H402, I62, and V62, and CFHR1-5. The final assay consisted of 24 peptides and 72 interference-free SRM transition ion pairs. Most peptides showed good linearity over 0.3–200 fmol/μL concentration range. Plasma concentrations of CFH variants and CFHR1-5 were measured using the SRM assay in 344 adults. Plasma CFH concentrations (mean, SE in μg/mL) by inferred genotype were: YY402, II62 (170.1, 31.4), YY402, VV62 (188.8, 38.5), HH402, VV62 (144.0, 37.0), HY402, VV62 (164.2, 42.3), YY402, IV62 (194.8, 36.8), HY402, IV62 (181.3, 44.7). Mean (SE) plasma concentrations of CFHR1-5 were 1.63 (0.04), 3.64 (1.20), 0.020 (0.001), 2.42 (0.18), and 5.49 (1.55) μg/mL, respectively. This SRM assay should facilitate the study of the role of systemic complement and risk of AMD.

KW - Age-related macular degeneration

KW - Biomedicine

KW - Complement factor H

KW - Complement factor H-related proteins

KW - Mass spectrometry

KW - Selected reaction monitoring

UR - http://www.scopus.com/inward/record.url?scp=85016213229&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85016213229&partnerID=8YFLogxK

U2 - 10.1002/pmic.201600237

DO - 10.1002/pmic.201600237

M3 - Article

C2 - 27647805

AN - SCOPUS:85016213229

VL - 17

JO - Proteomics

JF - Proteomics

SN - 1615-9853

IS - 6

M1 - 1600237

ER -