Imperial College London
Alternate

ProfessorMarkThursz

Faculty of Medicine, Department of Metabolism, Digestion and Reproduction

Professor of Hepatology. Head of Department
 
 
 
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Contact

 

+44 (0)20 3312 1903m.thursz

 
 
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Assistant

 

Ms Dawn Campbell +44 (0)20 3312 6454

 
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Location

 

Norfolk PlaceSt Mary's Campus

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Summary

 

Publications

Citation

BibTex format

@article{Kumar:2017:10.1038/s41598-017-12229-2,
author = {Kumar, A and Gangadharan, B and Cobbold, J and Thursz, M and Zitzmann, N},
doi = {10.1038/s41598-017-12229-2},
journal = {Scientific Reports},
title = {Absolute quantitation of disease protein biomarkers in a single LC-MS acquisition using apolipoprotein F as an example},
url = {http://dx.doi.org/10.1038/s41598-017-12229-2},
volume = {7},
year = {2017}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - LC-MS and immunoassay can detect protein biomarkers. Immunoassays are more commonly used but can potentially be outperformed by LC-MS. These techniques have limitations including the necessity to generate separate calibration curves for each biomarker. We present a rapid mass spectrometry-based assay utilising a universal calibration curve. For the first time we analyse clinical samples using the HeavyPeptide IGNIS kit which establishes a 6-point calibration curve and determines the biomarker concentration in a single LC-MS acquisition. IGNIS was tested using apolipoprotein F (APO-F), a potential biomarker for non-alcoholic fatty liver disease (NAFLD). Human serum and IGNIS prime peptides were digested and the IGNIS assay was used to quantify APO-F in clinical samples. Digestion of IGNIS prime peptides was optimised using trypsin and SMART Digest™. IGNIS was 9 times faster than the conventional LC-MS method for determining the concentration of APO-F in serum. APO-F decreased across NAFLD stages. Inter/intra-day variation and stability post sample preparation for one of the peptides was ≤13% coefficient of variation (CV). SMART Digest™ enabled complete digestion in 30 minutes compared to 24 hours using in-solution trypsin digestion. We have optimised the IGNIS kit to quantify APO-F as a NAFLD biomarker in serum using a single LC-MS acquisition.
AU  - Kumar,A
AU  - Gangadharan,B
AU  - Cobbold,J
AU  - Thursz,M
AU  - Zitzmann,N
DO  - 10.1038/s41598-017-12229-2
PY  - 2017///
SN  - 2045-2322
TI  - Absolute quantitation of disease protein biomarkers in a single LC-MS acquisition using apolipoprotein F as an example
T2  - Scientific Reports
UR  - http://dx.doi.org/10.1038/s41598-017-12229-2
UR  - http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000411416600028&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
UR  - http://hdl.handle.net/10044/1/54191
VL  - 7
ER  -
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