2016
DOI: 10.1007/s13361-016-1368-2
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Multiple Reaction Monitoring for Direct Quantitation of Intact Proteins Using a Triple Quadrupole Mass Spectrometer

Abstract: Abstract. Methods that can efficiently and effectively quantify proteins are needed to support increasing demand in many bioanalytical fields. Triple quadrupole mass spectrometry (QQQ-MS) is sensitive and specific, and it is routinely used to quantify small molecules. However, low resolution fragmentation-dependent MS detection can pose inherent difficulties for intact proteins. In this research, we investigated variables that affect protein and fragment ion signals to enable protein quantitation using QQQ-MS.… Show more

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Cited by 30 publications
(48 citation statements)
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“…Chromatogram of protein standards including PSA in the optimal separation condition established in this study with the re‐developed and optimized MRM method. The myoglobin signal is reduced by half to fit the scale .…”
Section: Resultsmentioning
confidence: 99%
See 1 more Smart Citation
“…Chromatogram of protein standards including PSA in the optimal separation condition established in this study with the re‐developed and optimized MRM method. The myoglobin signal is reduced by half to fit the scale .…”
Section: Resultsmentioning
confidence: 99%
“…The absence of alterations in the protein charge state distributions with different stationary phases echoes the results reported by Drake et al where Chromatogram of protein standards including PSA in the optimal separation condition established in this study with the re-developed and optimized MRM method. The myoglobin signal is reduced by half to fit the scale [19].…”
Section: Effects Of Mobile Phasementioning
confidence: 99%
“…On the other hand, top-down MS (TD-MS) identifies intact proteins and can detect sequence variants or provide a scaffold for sequencing, but it is about 100-fold less sensitive than BU-MS; it requires large (>7 to 14 T) magnets and generally lags BU-MS in terms of proteomic coverage and throughput ( Fig. 1B) (31,(54)(55)(56)(57)(58)(59)(60).TD-MS analysis introduces intact protein ions into the gas phase by electrospray ionization that are subsequently fragmented commonly by collisioninduced dissociation or, more gingerly, by electron-capture dissociation or electron-transfer dissociation in the mass spectrometer, yielding the masses of both the protein and the fragment ions. With enough fragments, this analysis can provide a comprehensive picture of the primary structure of the protein with its accompanying modifications.…”
Section: Beyond Genomics and Transcriptomics Toward Proteomics With Msmentioning
confidence: 99%
“…The LC/MS‐based chromatographic methodology developed used both formic acid and a small amount of trifluoroacetic acid in the mobile phase to yield high‐efficiency separations. As with the previous study, all of these studies focused on the analysis of relatively low‐molecular‐weight proteins.…”
Section: Introductionmentioning
confidence: 99%
“…Recently, Wang et al have demonstrated the use of triple quadrupole mass spectrometry in multiple reaction monitoring (MRM) mode for intact protein analysis . Their initial attempt to optimize conditions for top‐down quantitative analysis of low‐molecular‐weight proteins (9–28 kDa) was successful.…”
Section: Introductionmentioning
confidence: 99%