Mass spectrometry for the detection of differentially expressed proteins: a comparison of surface‐enhanced laser desorption/ionization and capillary electrophoresis/mass spectrometry

Neuhoff, Nils von; Kaiser, Thorsten; Wittke, Stefan; Krebs, Ronald; Pitt, Andrew R.; Burchard, Aurea; Sundmacher, Astrid; Schlegelberger, Brigitte; Kölch, Walter; Mischak, Harald

doi:10.1002/rcm.1294

Cited by 191 publications

(152 citation statements)

References 11 publications

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“…Mass spectral ion peaks representing identical molecules at different charge states were deconvoluted into single masses using MosaiquesVisu software [44]. Migration time and ion signal intensity (amplitude) were normalized using internal polypeptide standards [45], [46].…”

Section: Methodsmentioning

confidence: 99%

A Distinct Urinary Biomarker Pattern Characteristic of Female Fabry Patients That Mirrors Response to Enzyme Replacement Therapy

et al. 2011

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Female patients affected by Fabry disease, an X-linked lysosomal storage disorder, exhibit a wide spectrum of symptoms, which renders diagnosis, and treatment decisions challenging. No diagnostic test, other than sequencing of the alpha-galactosidase A gene, is available and no biomarker has been proven useful to screen for the disease, predict disease course and monitor response to enzyme replacement therapy. Here, we used urine proteomic analysis based on capillary electrophoresis coupled to mass spectrometry and identified a biomarker profile in adult female Fabry patients. Urine samples were taken from 35 treatment-naïve female Fabry patients and were compared to 89 age-matched healthy controls. We found a diagnostic biomarker pattern that exhibited 88.2% sensitivity and 97.8% specificity when tested in an independent validation cohort consisting of 17 treatment-naïve Fabry patients and 45 controls. The model remained highly specific when applied to additional control patients with a variety of other renal, metabolic and cardiovascular diseases. Several of the 64 identified diagnostic biomarkers showed correlations with measures of disease severity. Notably, most biomarkers responded to enzyme replacement therapy, and 8 of 11 treated patients scored negative for Fabry disease in the diagnostic model. In conclusion, we defined a urinary biomarker model that seems to be of diagnostic use for Fabry disease in female patients and may be used to monitor response to enzyme replacement therapy.

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Section: Methodsmentioning

confidence: 99%

A Distinct Urinary Biomarker Pattern Characteristic of Female Fabry Patients That Mirrors Response to Enzyme Replacement Therapy

et al. 2011

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“…CE-MS was compared to surface-enhanced laser desorption ionization (SELDI) MS [122]. SELDI profiles of polypeptides from urine of patients suffering from glomerulonephritis were sparser, showed lower resolution and mass accuracy, and also were much more concentration dependent than CE-MS. By comparing urine from patients and healthy control, SELDI-MS identified three polypeptides as potential markers for glomerulonephritis, whereas CE-MS found about 200 potential biomarkers among urinary polypeptides.…”

Section: Msmentioning

confidence: 99%

“…However, no single polypeptide was identified that would be characteristic for a specific disease and simultaneously absent in all control samples [207]. Urine of patients with kidney diseases provided 27 exclusive polypeptides and 13 others, present in control, were missing [122,212,215,216]. Similarly, CE-MS/MS was used to analyze urinary samples from 112 patients with type II diabetes.…”

Section: Urinary Proteinsmentioning

confidence: 99%

Capillary electrophoresis of proteins 2003–2005

Dolnı́k¹

2006

Electrophoresis

139

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This review article with 304 references describes recent developments in CE of proteins, and covers the two years since the previous review (Hutterer, K., Dolník, V., Electrophoresis 2003, 24, 3998-4012) through Spring 2005. It covers topics related to CE of proteins, including modeling of the electrophoretic migration of proteins, sample pretreatment, wall coatings, improving separation, various forms of detection, special electrophoretic techniques such as affinity CE, CIEF, and applications of CE to the analysis of proteins in real-world samples including human body fluids, food and agricultural samples, protein pharmaceuticals, and recombinant protein preparations.

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“…The essential information that needs to be extracted from an entire CE-MS run is the identity and quantity of detected polypeptides. We therefore developed MosaiquesVisu (available at www.proteomiques.com) as an approach towards extracting information from these highly complex spectra [12].…”

Section: Software and Data Analysismentioning

confidence: 99%

“…This was certainly a major step forward with respect to practicability, since alternative approaches like 2-DE followed by MS or HPLC-MS could be operated only by skilled personnel and were time consuming. Unfortunately, soon the price for simplicity became obvious: only a very small fraction of polypeptides was detectable on the chip surface [12]. The low resolution and accuracy -while easing interpretation of the datatogether with debatable study design raised serious concerns about the validity of the data [13][14][15].…”

Section: Introductionmentioning

confidence: 99%

Capillary electrophoresis coupled to mass spectrometry for clinical diagnostic purposes

2005

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Capillary electrophoresis coupled to mass spectrometry for clinical diagnostic purposesThe introduction of fast, sensitive, and robust techniques for proteomic analysis into clinical practice represents a major step toward a new diagnostic approach of body fluids. In addition, proteomics emerges as a key technology for the discovery of disease biomarkers in various body fluids. However, even in relatively protein-deprived body fluids such as urine, the complexity and wide dynamic range of protein expression pose a considerable challenge to both separation and identification technologies. In the present review we discuss from a clinical point-of-view recent advances of the use of proteomics in clinical diagnosis as well as therapy evaluation. We focus on capillary electrophoresis coupled to mass spectrometry (CE-MS) and discuss CE-MS from an application point of view evaluating its merits and vices with regard to biomarker discovery. This review further presents examples of clinical applications of CE-MS for detection and identification of biomarkers in urine. IntroductionIt is well appreciated for centuries that body fluids contain information on the "internal" environment, i.e., the function of tissues and organs, which can be used as a gauge for the well being of the organism. Although this concept was created by Hippocrates more than 2000 years ago, standardized clinical laboratory tests were not developed before the 20th century. Moreover, a systematic discovery of biomarkers was made possible only with the discovery of antibodies and their use in immunological assays such as radioimmuno assay (RIA), ELISA, etc. Their increasing use enabled detection and quantification of distinct molecules -mainly peptides and proteins -in various body fluids. However, these techniques were limited by their requirement of prior knowledge of the molecule and a specific probe. New functional proteomic approaches appeared to circumvent these limitations. Spurred by advances in MS it was hoped that proteomics would allow the analysis of thousands of peptides and proteins in one sample [1][2][3][4][5][6], but several obstacles turned up soon. The high complexity and wide dynamic range of peptides in body fluids required pre-MS separation techniques that offer high resolution and could cope with a wide dynamic range of peptide concentrations [3]. Further, suitable software for interpretation of a vast number of generated MS spectra as well as comparison of the data was a major challenge [7].The approach towards solving most of these problems was to reduce data complexity, for example by arbitrarily collecting data on only a few polypeptides and also to reduce resolution of the analysis instrument. Both were accomplished with the surface-enhanced laser desorption/ionization (SELDI) technology -a complex solution of analytes was allowed to interact with an active surface (ion-exchange, hydro-

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Mass spectrometry for the detection of differentially expressed proteins: a comparison of surface‐enhanced laser desorption/ionization and capillary electrophoresis/mass spectrometry

Cited by 191 publications

References 11 publications

A Distinct Urinary Biomarker Pattern Characteristic of Female Fabry Patients That Mirrors Response to Enzyme Replacement Therapy

A Distinct Urinary Biomarker Pattern Characteristic of Female Fabry Patients That Mirrors Response to Enzyme Replacement Therapy

Capillary electrophoresis of proteins 2003–2005

Capillary electrophoresis coupled to mass spectrometry for clinical diagnostic purposes

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