UCL Great Ormond Street Institute of Child Health


Great Ormond Street Institute of Child Health


Targeted Proteomics

Protein quantitation by mass spectrometry

Targeted proteomics or Multiple Reaction Monitoring (MRM) LC-MS/MS analysis is a powerful technique that can multiplex the quantitation of multiple proteins in one analysis.

Essentially a protein or biomarker can be measured in a complex sample such as serum, CSF or urine by identifying a unique peptide sequence (proteotypic) that represents that protein. The sample is then digested using a protease to generate that peptide from the biomarker protein. The unique peptide is then measured using mass spectrometry. The mass spectrometer can measure many peptides at once so an assay can be ‘multiplexed’


Targeted proteomics schematic


The TMSRG has extensive experience developing multiplex targeted proteomic assays

We have assays developed in CSF, urine serum/plasma to confirm biomarker or measure the inflammation profile. 

We are now using these assays to look at the effect of  post –covid19 infection on healthcare workers in collaboration with Great Ormond Street, Barts and the Royal Free Hospitals. 


Please see below for a list of our publications using targeted proteomics:


Ambroxol for the Treatment of Patients With Parkinson Disease With and Without Glucocerebrosidase Gene Mutations: A Nonrandomized, Noncontrolled Trial.

Mullin S, Smith L, Lee K, D'Souza G, Woodgate P, Elflein J, Hällqvist J, Toffoli M, Streeter A, Hosking J, Heywood WE, Khengar R, Campbell P, Hehir J, Cable S, Mills K, Zetterberg H, Limousin P, Libri V, Foltynie T, Schapira AHV.

JAMA Neurol. 2020 Apr 1;77(4):427-434. doi: 10.1001/jamaneurol.2019.4611.

Rapid, proteomic urine assay for monitoring progressive organ disease in Fabry disease.

Doykov ID, Heywood WE, Nikolaenko V, Śpiewak J, Hällqvist J, Clayton PT, Mills P, Warnock DG, Nowak A, Mills K.

J Med Genet. 2020 Jan;57(1):38-47. doi: 10.1136/jmedgenet-2019-106030. Epub 2019 Sep 13.

Identification of a Multiplex Biomarker Panel for Hypertrophic Cardiomyopathy Using Quantitative Proteomics and Machine Learning.

Captur G, Heywood WE, Coats C, Rosmini S, Patel V, Lopes LR, Collis R, Patel N, Syrris P, Bassett P, O'Brien B, Moon JC, Elliott PM, Mills K.

Mol Cell Proteomics. 2020 Jan;19(1):114-127. doi: 10.1074/mcp.RA119.001586. Epub 2019 Jun 26.

Preparation of iPSCs for Targeted Proteomic Analysis.

Baud A, Heywood WE, Little D, Gissen P, Mills K.

Methods Mol Biol. 2019;1994:131-139. doi: 10.1007/978-1-4939-9477-9_11.

CSF pro-orexin and amyloid-β38 expression in Alzheimer's disease and frontotemporal dementia.

Heywood WE, Hallqvist J, Heslegrave AJ, Zetterberg H, Fenoglio C, Scarpini E, Rohrer JD, Galimberti D, Mills K.

Neurobiol Aging. 2018 Dec;72:171-176. doi: 10.1016/j.neurobiolaging.2018.08.019. Epub 2018 Aug 25.

Multiplex High-Throughput Targeted Proteomic Assay To Identify Induced Pluripotent Stem Cells.

Baud A, Wessely F, Mazzacuva F, McCormick J, Camuzeaux S, Heywood WE, Little D, Vowles J, Tuefferd M, Mosaku O, Lako M, Armstrong L, Webber C, Cader MZ, Peeters P, Gissen P, Cowley SA, Mills K.

Anal Chem. 2017 Feb 21;89(4):2440-2448. doi: 10.1021/acs.analchem.6b04368. Epub 2017 Feb 6.

A targeted proteomic multiplex CSF assay identifies increased malate dehydrogenase and other neurodegenerative biomarkers in individuals with Alzheimer's disease pathology.

Paterson RW, Heywood WE, Heslegrave AJ, Magdalinou NK, Andreasson U, Sirka E, Bliss E, Slattery CF, Toombs J, Svensson J, Johansson P, Fox NC, Zetterberg H, Mills K, Schott JM.

Transl Psychiatry. 2016 Nov 15;6(11):e952. doi: 10.1038/tp.2016.194.

A High Throughput, Multiplexed and Targeted Proteomic CSF Assay to Quantify Neurodegenerative Biomarkers and Apolipoprotein E Isoforms Status.

Heywood WE, Baud A, Bliss E, Sirka E, Schott JM, Zetterberg H, Galimberti D, Sebire NJ, Mills K.

J Vis Exp. 2016 Oct 20;(116):54541. doi: 10.3791/54541.

Increased cerebrospinal fluid soluble TREM2 concentration in Alzheimer's disease.

Heslegrave A, Heywood W, Paterson R, Magdalinou N, Svensson J, Johansson P, Öhrfelt A, Blennow K, Hardy J, Schott J, Mills K, Zetterberg H.

Mol Neurodegener. 2016 Jan 12;11:3. doi: 10.1186/s13024-016-0071-x.

Identification of novel CSF biomarkers for neurodegeneration and their validation by a high-throughput multiplexed targeted proteomic assay.

Heywood WE, Galimberti D, Bliss E, Sirka E, Paterson RW, Magdalinou NK, Carecchio M, Reid E, Heslegrave A, Fenoglio C, Scarpini E, Schott JM, Fox NC, Hardy J, Bhatia K, Heales S, Sebire NJ, Zetterberg H, Mills K.

Mol Neurodegener. 2015 Dec 1;10:64. doi: 10.1186/s13024-015-0059-y.

Proteomic Discovery and Development of a Multiplexed Targeted MRM-LC-MS/MS Assay for Urine Biomarkers of Extracellular Matrix Disruption in Mucopolysaccharidoses I, II, and VI.

Heywood WE, Camuzeaux S, Doykov I, Patel N, Preece RL, Footitt E, Cleary M, Clayton P, Grunewald S, Abulhoul L, Chakrapani A, Sebire NJ, Hindmarsh P, de Koning TJ, Heales S, Burke D, Gissen P, Mills K.

Anal Chem. 2015 Dec 15;87(24):12238-44. doi: 10.1021/acs.analchem.5b03232. Epub 2015 Nov 20.

The identification of new biomarkers for identifying and monitoring kidney disease and their translation into a rapid mass spectrometry-based test: evidence of presymptomatic kidney disease in pediatric Fabry and type-I diabetic patients.

Manwaring V, Heywood WE, Clayton R, Lachmann RH, Keutzer J, Hindmarsh P, Winchester B, Heales S, Mills K.

J Proteome Res. 2013 May 3;12(5):2013-21. doi: 10.1021/pr301200e. Epub 2013 Apr 2.