Liquid MALDI MS Analysis of Complex Peptide and Proteome Samples

Wiangnon, Kanjana; Cramer, Rainer

doi:10.1021/acs.jproteome.6b00148

Cited by 8 publications

(6 citation statements)

References 27 publications

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“…Without enrichment (Fig. 5C), the sample contains mostly peptide fragments below m / z 1500, as woud be expected from a BSA digest . The presence of the higher‐mass peptides characteristic of a β‐casein digest, which are present in the tryptic digest mixture at the picomol level (∼0.4 pmol), are barely noticeable.…”

Section: Resultsmentioning

confidence: 75%

Application of polydopamine‐coated nylon capillary‐channeled polymer fibers as a stationary phase for mass spectrometric phosphopeptide analysis

Trang

Marcus

2019

Electrophoresis

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Application of polydopamine-coated nylon capillary-channeled polymer fibers as a stationary phase for mass spectrometric phosphopeptide analysisCapillary-channeled polymer (C-CP) fibers are demonstrated as a selective stationary phase for phosphopeptide analysis via LC-MS. Taking advantage of the oxidative selfpolymerization of dopamine under alkaline conditions, a simple system involving a dilute aqueous solution of 0.2% w/v dopamine hydrochloride in 0.15% w/v TRIS buffer, pH 8.5 was utilized to coat polydopamine onto nylon 6 C-CP fibers. Confirmation of the polydopamine coating on the fibers (nylon-PDA) was made through attenuated total reflection-FTIR (ATR-FTIR) analysis. Imaging using SEM was also performed to examine the morphology and topography of the nylon-PDA. Subsequent loading of Fe 3+ to the nylon-PDA matrix was confirmed by SEM/energy dispersive X-ray spectroscopy (SEM/EDX). The Fe 3+ -bound nylon-PDA fibers packed in a microbore column format were tested in the offline preconcentration of phosphopeptides from a 1:100 mixture of β-casein/BSA digests for MALDI-TOF analysis. The packed column was also installed onto an HPLC system as a platform for the online sample clean-up and enrichment of phosphopeptides from a 1:1000 mixture of β-casein/BSA protein digests that were determined by subsequent ESI-MS analysis.Abbreviations: ATR-FTIR, attenuated total reflection-FTIR; C-CP, capillary-channeled polymer; EDX, energy-dispersive X-ray spectroscopy; IMAC, immobilized metal affinity chromatography; IDA, iminodiacetic acid; MOAC, metal oxide affinity chromatography; NTA, nitriloacetic acid; PDA, polydopamine chiometry of protein phosphorylation [4]. In addition, phosphopeptides are known for their poor ionization efficiencies due to their associated negative charge. Moreover, the low abundance of phosphopeptides in complex peptide mixtures is also a challenge. In order to overcome these two technical difficulties, use of some form of sample clean-up is critically required to isolate and pre-concentrate phosphopeptides from complex peptide matrices [5].Among the different approaches of selective enrichment of phosphopeptides are immobilized metal ion affinity chromatography (IMAC) and metal oxide affinity chromatography (MOAC) [6]. IMAC utilizes the affinity of the phosphate groups in phosphopeptides to metal ions (Fe 3+ or Ga 3+ ) immobilized on the stationary phase via the acidic chelating ligands of iminodiacetic acid (IDA) or nitriloacetic acid (NTA) [7,8]. The conventional IMAC technique has two main disadvantages. First, it experiences possible leakage of metals during sample loading and washing steps. Second, the selectivity of the traditional IMAC adsorbents (which is typically low) is interfered by the non-phosphopeptides containing acidic amino acid residues (such as glutamate and aspartate) [8,9]. The latter can be partially addressed by methyl esterification of carboxylic acid groups of peptides; however,

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Section: Resultsmentioning

confidence: 75%

Application of polydopamine‐coated nylon capillary‐channeled polymer fibers as a stationary phase for mass spectrometric phosphopeptide analysis

Trang

Marcus

2019

Electrophoresis

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“…All analyte solutions were analysed by liquid AP-MALDI MS 10,21,22 . Briefly, liquid MALDI samples were prepared using α-cyano-4-hydroxycinnamic acid (CHCA) as the MALDI matrix chromophore.…”

Section: Methodsmentioning

confidence: 99%

Speciation and milk adulteration analysis by rapid ambient liquid MALDI mass spectrometry profiling using machine learning

Piras

Hale

Reynolds

et al. 2021

Sci Rep

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Growing interest in food quality and traceability by regulators as well as consumers demands advances in more rapid, versatile and cost-effective analytical methods. Milk, as most food matrices, is a heterogeneous mixture composed of metabolites, lipids and proteins. One of the major challenges is to have simultaneous, quantitative detection (profiling) of this panel of biomolecules to gather valuable information for assessing food quality, traceability and safety. Here, for milk analysis, atmospheric pressure matrix-assisted laser desorption/ionization employing homogenous liquid sample droplets was used on a Q-TOF mass analyzer. This method has the capability to produce multiply charged proteinaceous ions as well as highly informative profiles of singly charged lipids/metabolites. In two examples, this method is coupled with user-friendly machine-learning software. First, rapid speciation of milk (cow, goat, sheep and camel) is demonstrated with 100% classification accuracy. Second, the detection of cow milk as adulterant in goat milk is shown at concentrations as low as 5% with 92.5% sensitivity and 94.5% specificity.

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“…The combined lipid and protein MS analyses were performed by liquid AP-MALDI MS [ 21 , 22 , 23 ] on a Synapt G2-Si High Definition Mass Spectrometry (HDMS) mass spectrometer (Waters Corporation) equipped with an in-house developed AP-MALDI source, using a Waters research enabled software (WREnS)-controlled XY stage (Zaber Technologies Inc., Vancouver, Canada). A detailed source description has been reported previously [ 24 ].…”

Section: Methodsmentioning

confidence: 99%

Rapid Liquid AP-MALDI MS Profiling of Lipids and Proteins from Goat and Sheep Milk for Speciation and Colostrum Analysis

et al. 2020

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Rapid profiling of the biomolecular components of milk can be useful for food quality assessment and for food fraud detection. Differences in commercial value and availability of milk from specific species are often the reasons for the illicit and fraudulent sale of milk whose species origin is wrongly declared. In this study, a fast, MS-based speciation method is presented to distinguish sheep from goat milk and sheep colostrum at different phases. Using liquid atmospheric pressure (AP)-matrix-assisted laser desorption/ionisation (MALDI) MS, it was possible to classify samples of goat and sheep milk with 100% accuracy in one minute of data acquisition per sample. Moreover, an accuracy of 98% was achieved in classifying pure sheep milk samples and sheep milk samples containing 10% goat milk. Evaluating colostrum quality and postnatal stages represents another possible application of this technology. Classification of sheep colostrum samples that were collected within 6 hours after parturition and 48 hours later was achieved with an accuracy of 84.4%. Our data show that substantial changes in the lipid profile can account for the accurate classification of colostrum collected at the early and late time points. This method applied to the analysis of protein orthologs of different species can, as in this case, allow unequivocal speciation analysis.

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Liquid MALDI MS Analysis of Complex Peptide and Proteome Samples

Cited by 8 publications

References 27 publications

Application of polydopamine‐coated nylon capillary‐channeled polymer fibers as a stationary phase for mass spectrometric phosphopeptide analysis

Application of polydopamine‐coated nylon capillary‐channeled polymer fibers as a stationary phase for mass spectrometric phosphopeptide analysis

Speciation and milk adulteration analysis by rapid ambient liquid MALDI mass spectrometry profiling using machine learning

Rapid Liquid AP-MALDI MS Profiling of Lipids and Proteins from Goat and Sheep Milk for Speciation and Colostrum Analysis

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