Multisegment injections improve peptide identification rates in capillary zone electrophoresis-based bottom-up proteomics

Boley, Danielle A.; Zhang, Zhenbin; Dovic̀hi, Norman J.

doi:10.1016/j.chroma.2017.07.022

Cited by 28 publications

(25 citation statements)

References 28 publications

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“…Boley et al. showed a doubling in the peptide identification rate . Using such an approach in an offline two‐dimensional RPLC‐CZE‐MS system, Faserl et al.…”

Section: Analytical Applicationsmentioning

confidence: 99%

“…Sequential injection of samples can increase the throughput of CE-MS by lowering dead times between runs. Boley et al showed a doubling in the peptide identification rate [58]. Using such an approach in an offline two-dimensional RPLC-CZE-MS system, Faserl et al identified 62 000 peptides, representing 6100 proteins in a 12.5 h run by sequential injection of RPLC fractions every 10 min [59].…”

Section: Bottom-up Proteomicsmentioning

confidence: 99%

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Recent advances in capillary electrophoresis‐mass spectrometry: Instrumentation, methodology and applications

et al. 2018

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Capillary electrophoresis (CE) offers fast and high‐resolution separation of charged analytes from small injection volumes. Coupled to mass spectrometry (MS), it represents a powerful analytical technique providing (exact) mass information and enables molecular characterization based on fragmentation. Although hyphenation of CE and MS is not straightforward, much emphasis has been placed on enabling efficient ionization and user‐friendly coupling. Though several interfaces are now commercially available, research on more efficient and robust interfacing with nano‐electrospray ionization (ESI), matrix‐assisted laser desorption/ionization (MALDI) and inductively coupled plasma mass spectrometry (ICP) continues with considerable results. At the same time, CE‐MS has been used in many fields, predominantly for the analysis of proteins, peptides and metabolites. This review belongs to a series of regularly published articles, summarizing 248 articles covering the time between June 2016 and May 2018. Latest developments on hyphenation of CE with MS as well as instrumental developments such as two‐dimensional separation systems with MS detection are mentioned. Furthermore, applications of various CE‐modes including capillary zone electrophoresis (CZE), nonaqueous capillary electrophoresis (NACE), capillary gel electrophoresis (CGE) and capillary isoelectric focusing (CIEF) coupled to MS in biological, pharmaceutical and environmental research are summarized.

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“…Boley et al. showed a doubling in the peptide identification rate . Using such an approach in an offline two‐dimensional RPLC‐CZE‐MS system, Faserl et al.…”

Section: Analytical Applicationsmentioning

confidence: 99%

Section: Bottom-up Proteomicsmentioning

confidence: 99%

Recent advances in capillary electrophoresis‐mass spectrometry: Instrumentation, methodology and applications

et al. 2018

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“…Thereafter, it was developed to solve the problem of lacking IS in protein analysis via successive injections of the standard protein at a known concentration followed by the sample to be quantified . Along with the successful applications in metabolic profiling and bottom‐up proteomics , MSI–CE–MS has been accepted as a very efficient approach for increasing the throughput of metabolomics and proteomics analysis .…”

Section: Introductionmentioning

confidence: 99%

Application of multisegment injection on quantification of creatinine and standard addition analysis of urinary 5‐hydroxyindoleacetic acid simultaneously with creatinine normalization

Huang

Scotland

et al. 2020

Electrophoresis

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Application of multisegment injection on quantification of creatinine and standard addition analysis of urinary 5-hydroxyindoleacetic acid simultaneously with creatinine normalizationIn this paper, the development of a simple dilute-and-shoot method for quantifying urinary creatinine by CE-ESI-MS was described. The creatinine analysis time was about 7 min/sample by conventional single injection (SI) method and can be significantly reduced to less than 2 min/sample with multi-segment injection (MSI). In addition, the standard addition analysis of 5-hydroxyindole-3-acetic acid (5-HIAA) and creatinine normalization was performed within one run by the MSI technique, and the total analysis time was 14-min faster compared to the SI method for analyzing the same set of samples. The uses of isotopic and non-isotopic internal standards (ISs) were compared. Creatinine-(methyl-13 C) and 5-hydroxyindole-4,6,7-D 3 -3-acetic-D 2 acid (5-HIAA-D 5 ) used as isotopic ISs can provide both accurate and precise results. In contrast, 1,5,5-trimethylhydantoin (1,5,5-TH) used as the non-isotopic IS for creatinine may cause a bias of over 13% in SI method and even worse when the MSI technique was used. Another compound, 2-methyl-3-indoleacetic acid (2-MIAA), was determined not suitable for MSI analysis of 5-HIAA due to endogenous interferences despite its acceptable performance in conventional methods of analysis.Abbreviations: 1,5,5-TH, 1,5,5-trimethylhydantoin; 2-MIAA, 2methyl-3-indoleacetic acid; 5-HIAA, 5-hydroxyindole-3-acetic acid; IS, internal standard; MRM, multiple reaction monitoring; MSI, multi-segment injection; RPA, relative peak area; SI, single injection variations due to water intake or diet composition. This gives rise to the difficulty of interpreting urinary metabolite concentrations and necessitating urine volume calibration [2,3].The conventional urine normalization approaches include flow rate correction, measurement of urine osmolality or specific gravity, and the commonly used creatinine correction [2,3]. The daily renal excretion of creatinine via glomerular filtration remains at a relatively constant rate under normal conditions [2-4], and the urinary creatinine level has been found relatively stable when the urine secretion rates are above 1 mL/min [3,5]. Despite the potential influence of age, gender, diet intake, and other factors, creatinine normalization is still widely applied and generally accepted in metabolomic studies for correcting solute concentrations of urine samples [1][2][3]. Furthermore, urinary creatinine levels are also measured for the assessment of renal function in Color online: See the article online to view Figs. 1-4 in color.

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“…Very recently Boley et al. performed three sequential injections of a yeast proteome digest . However, since both groups injected the same sample multiple times the increase in proteome coverage is limited.…”

mentioning

confidence: 99%

“…[14] Very recently Boley et al performed three sequential injections of a yeast proteome digest. [15] However, since both groups injected the same sample multiple times the increase in proteome coverage is limited. To evaluate the sequential injection CE-MS method for its applicability in proteomics research we extracted and digested whole proteins from human lymphoblastic T-cells.…”

mentioning

confidence: 99%

Enhancing Proteomic Throughput in Capillary Electrophoresis–Mass Spectrometry by Sequential Sample Injection

et al. 2017

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In this study we demonstrate the potential of sequential injection of samples in capillary electrophoresis-mass spectrometry for rapid and sensitive proteome characterization of human lymphoblastic T-cells (line CCRF-CEM). Proteins were extracted, enzymatically digested, and the resulting peptides fractionated by RP-HPLC. Twenty fractions were thereafter analyzed by CE-MS within a single MS analysis. The CE-MS method was designed so that every 10 min a new fraction was injected into the CE system. Without any rinsing or equilibration steps we were able to generate a continuous stream of peptides feeding the mass analyzer. In 250 min, the total analysis time of a single sequential injection experiment, we were able to identify roughly 28 000 peptide sequences counting for 4800 proteins. These numbers could be increased to 62 000 peptides and more than 6100 proteins identified, when performing three experiments analyzing a total of 60 fractions, all within 12.5 h. We found that the electrophoretic mobility of peptides can be used to trace back peptides and assign them to the fraction they originate from.

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Multisegment injections improve peptide identification rates in capillary zone electrophoresis-based bottom-up proteomics

Cited by 28 publications

References 28 publications

Recent advances in capillary electrophoresis‐mass spectrometry: Instrumentation, methodology and applications

Recent advances in capillary electrophoresis‐mass spectrometry: Instrumentation, methodology and applications

Application of multisegment injection on quantification of creatinine and standard addition analysis of urinary 5‐hydroxyindoleacetic acid simultaneously with creatinine normalization

Enhancing Proteomic Throughput in Capillary Electrophoresis–Mass Spectrometry by Sequential Sample Injection

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