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Download fileMetal-Assisted Protein Quantitation (MAPq): Multiplex Analysis of Protein Expression Using Lanthanide-Modified Antibodies with Detection by Inductively Coupled Plasma Mass Spectrometry
journal contribution
posted on 18.05.2020, 18:03 authored by Sarah
C. Shuck, Cu Nguyen, Yin Chan, Timothy O’Connor, Alexandra K. Ciminera, Michael Kahn, John TerminiUnderstanding
the complex relationships between genomics, transcriptomics,
and proteomics requires the development of more sensitive and rapid
methods of multiplexed protein analysis. This is necessary to understand
the relationship between cellular responses to environmental stresses,
disease progression, and/or drug treatment; however, most methods
are limited by low sensitivity, nonspecificity, and minimal multiplexing
capacity. To more fully explore the relationship between multiple
cellular pathways, we have developed a novel antibody-based multiplex
assay using inductively coupled plasma mass spectrometry (ICP-MS),
which we term metal-assisted protein quantitation (MAPq). MAPq utilizes
lanthanide-conjugated antibodies to simultaneously quantify up to
35 proteins with low pg/mL sensitivity. This method is especially
advantageous for low-abundance proteins, a significant limitation
of many multiplex MS methods. We observed a limit of detection of
0.5 pg/mL and a limit of quantitation of 5 pg/mL with virtually no
background signal. We applied this method to both cultured cells and
mouse tissues to investigate changes in low-abundance nuclear and
cytoplasmic proteins following drug or environmental stresses. MAPq
was found to be at least 10 times more sensitive than Western blots
and could detect quantitative changes in protein expression not readily
observed using conventional approaches.
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Keywords
multiplex MS methodsmultiplexed protein analysisbackground signalLanthanide-Modified Antibodiescytoplasmic proteinsMetal-Assisted Protein Quantitation35 proteinsWestern blotsplasma mass spectrometryterm metal-assisted protein quantitationpgrelationship10 timesmultiplexing capacityPlasma Mass Spectrometrylanthanide-conjugated antibodiesProtein Expressionlow-abundance proteinsmouse tissuesMAPqmultiplex assayprotein expressiondisease progressionICP-MS