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 D.
N1 - Funding Information:
This work was supported by the National Institutes of Health grants R01 EY024596, R01 AG027012, R56 AG052973, the Intramural Research Program of the National Institute on Aging, the Joint King Khaled Eye Specialist Hospital and Wilmer Eye Institute Research Grant Program, the Edward N. & Della L. Thome Memorial Foundation, and Research to Prevent Blindness. The AGES Reykjavik study was supported by National Institutes of Health (contract nos. N01-AG-12100 and HHSN271201200022C); the National Institute on Aging Intramural Research Program; Hjartavernd (the Icelandic Heart Association); the Althingi (the Icelandic Parliament). [Corrections were made to this article on 21 November 2016, after acceptance: in Table, Figures and and Results, paragraphs 2 and 6.]
Publisher Copyright:
© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
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
SN - 1615-9853
VL - 17
JO - Proteomics
JF - Proteomics
IS - 6
M1 - 1600237
ER -