MRM

Multiple Reaction Monitoring (MRM, also referred to as SRM) is a tandem MS (MS/MS) scan mode unique to triple quadrupole MS instrumentation that monitors generation of a fragment ion by collision induced dissociation (CID) from an isolated precursor ion. MRM is a highly sensitive and targeted approach that requires knowledge of the molecular weight of an analyte and its fragmentation behavior under CID.

MRM is capable of accurate quantitation of numerous proteins in complex systems within a single experiment and is therefore ideal for biomarker validation and biomarker discovery. Although (MRM) assays have been widely employed for many years in pharmaceutical companies to quantify small molecules in biological specimens, its applicability towards quantitation of peptides for e.g. in blood samples has just been demonstrated recently. In particular, the Centre’s research is focused on the development of MRM approaches that use isotopically-coded peptides as standards enabling absolute quantitation of multiple proteins. As part of the Centre’s research program, cocktails of these standards are currently being created for profiling hundreds of proteins including blood proteins linked to cardiovascular diseases (CVD); for examining the endocrine pathways which can be disrupted by environmental perturbation; and for quantifying enzymes involved in the defense mechanism of trees against microbial attack. The specific proteins that we are targeting in the project of elucidating the defense mechanism are functionally different but structurally very similar and therefore difficult to quantify by the more-common ELISA technique. The peptide cocktails developed for the CVD project will be employed in large-scale proteomic analyses. As can be seen from these few examples, these “cocktails” have great potential for a wide range of applications.

Standard peptides are available through the Proteomics Centre (please visit Peptide Synthesis for additional information), and peptide cocktails and MRM methods currently developed are available by contacting the Centre directly. Furthermore, the Centre offers the development and implementation of complete and customized MRM assays as well as conducting these assays.

Selected Proteomics Centre MRM Publications
  1. Kuzyk, M.A., Smith, D., Yang, J., Cross T., Jackson, A., Hardie, D., , Anderson, N.L., Borchers, C.H. MRM-based, Multiplexed, Absolute Quantitation of 45 Proteins in Human Plasma. Mol and Cell Proteomics, 8(8):1860-77. (2009)

Human plasma proteins (73) for which we have developed MRM assays:

Adiponectin
Afamin
Albumin, serum
Aldolase C
Alpha-1-acid glycoprotein 1
Alpha-1-antichymotrypsin
Alpha-1-Anti-trypsin
Alpha-1B-glycoprotein
Alpha-2-antiplasmin
alpha-2-HS-glycoprotein
alpha-2-macroglobulin
Angiotensinogen
Antithrombin-III
Apolipoprotein A-I
Apolipoprotein A-II precursor
Apolipoprotein A-IV
Apolipoprotein B-100
Apolipoprotein C-I lipoprotein
Apolipoprotein C-II
Apolipoprotein C-III
Apolipoprotein D
Apolipoprotein E
Apolipoprotein L1
Apolipoprotein(a)
Aspartate aminotransferase,mitochondrial (m-type)
Beta-2-glycoprotein I
CD105-endoglin
Ceruloplasmin
Clusterin
Coag. Factor X
Coagulation Factor VII-activating Protease
Coagulation factor XIIa HC
Coagulation Factor XIII (a chain)
Coagulation Factor XIII B chain
Complement C1 inactivator
Complement C3
Complement C4 beta chain
Complement C4 gamma chain
Complement factor B
Complement factor H
ComplementC9
CRP
Fibrinogen alpha chain
Fibrinogen beta chain
Fibrinogen gamma chain
Fibrinopeptide A
Fibronectin
Gelsolin, isoform 1
Glial fibrillary acidic protein (GFAP)
Haptoglobin beta chain
Hemopexin
Heparin cofactor II
Histidine-rich glycoprotein
IGF-1
Inter-alpha-trypsin inhibitor HC
Kininogen-1
L-selectin
NGAL
Paraoxonase (PON 1) Peptide Synthesis
Plasma retinol-binding protein
Plasminogen
Protein C inhibitor
Protein S
Prothrombin
Serum amyloid P-component
Thrombospondin-1
Transferrin
Transthyretin
Tropomyosin 1 alpha chain
Vitamin D-binding protein
Vitronectin
von Willebrand Factor
Zinc-alpha-2-glycoprotein

Mouse plasma proteins (35) for which we have developed MRM assays:

Adiponectin
Aldolase C
Antithrombin-III
APO4
Apolipoprotein E
Aspartate aminotransferase,mitochondrial (m-type)
Catalase
Ceruloplasmin
Coagulation Factor V
Coagulation Factor VII-activating protease
Coagulation Factor X
Coagulation Factor XIII B chain
Complement C1 inactivator
creatine kinase-MB M chain
Erythropoeitin
Erythropoietin
Ferritin H
Ferritin L
Fibronectin
Haptoglobin
Haptoglobin beta chain
Hemojuvelin
Hemopexin
HO-1
Lactoferrin
matrix metalloproteinase-2 (MMP-2)
MMP8
natriuretic peptide, atrial, C-terminal (C-ANP)
Paraoxonase (PON 1)
protein C
Thrombospondin-1
tissue inhibitor of metalloproteinases-2 (TIMP-2)
Transferrin
Transferrin receptor 1
von Willebrand Factor