Influences of Blood Sample Processing on Low–Molecular-Weight Proteome Identified by Surface-Enhanced Laser Desorption/Ionization Mass Spectrometry

Banks, Rosamonde E.; Stanley, Anthea J.; Cairns, David A.; Barrett, Jennifer; Clarke, Paul; Thompson, Douglas; Selby, Peter J.

doi:10.1373/clinchem.2005.051417

Cited by 215 publications

(151 citation statements)

References 46 publications

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“…The CSF normal protein concentration is in the range of 20-40 mg per 100 mL in the lumbar cistern level [2], a value 100-times lower than in the blood. It is considered that 80% of this protein content derives from blood (through the choroid tissues) and 20% originates directly from the CNS [3]. The major proteins in both fluids are found similar except for high molecular weight proteins such as multimeric IgM that are not well transported through the blood-brain barrier.…”

Section: Cerebrospinal Fluid (Csf) Origin and Global Compositionmentioning

confidence: 99%

Clinical proteomics of the cerebrospinal fluid: Towards the discovery of new biomarkers

Roche

Gabelle

Lehmann

2008

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The cerebrospinal fluid (CSF) circulates within the CNS where it plays an essential physiological role in homeostasis of neuronal cells. This biological fluid has an important protein diversity that results from both filtration of serum through the blood-brain barrier and production/secretion of neuronal peptides and proteins. Changes in CSF composition depend on blood proteome, CSF circulation alterations, as well as physiological or pathological brain status. Hence, CSF proteomic analysis gives a unique opportunity to detect and describe biomarkers in neurological affections. Although lumbar puncture is considered as invasive, post lumbar puncture events remain rare and minor. Nevertheless, CSF biological analysis is currently limited to a small number of parameters and clinical situations suggesting mainly meningitis, malignancies or dementia. Few CSF proteomic studies have been performed in comparison to those on blood. In this review, we will provide a proteomics description of the CSF, summarize the current clinical use of this fluid and describe its clinical proteomics examination.

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Section: Cerebrospinal Fluid (Csf) Origin and Global Compositionmentioning

confidence: 99%

Clinical proteomics of the cerebrospinal fluid: Towards the discovery of new biomarkers

Roche

Gabelle

Lehmann

2008

Proteomics Clinical Apps

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“…Despite this, proteomic coverage is still limited and few if any robust cancer biomarkers have been identified using classical proteomic profiling methods. The need for identical handling of clinical samples is also of paramount importance in order to avoid largely proteolysis‐driven changes unrelated to biological variation and has been highlighted by numerous studies 12, 13, 14, 15, 16, 17. Finally, issues related to reproducibility and the robustness of class‐discriminating algorithms used for proteomic biomarker discovery have also been raised 18, 19.…”

Section: Introductionmentioning

confidence: 99%

Discovery of serum biomarkers of ovarian cancer using complementary proteomic profiling strategies

Timms

Arslan-Low

Kabir

et al. 2014

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PurposeOvarian cancer is a devastating disease and biomarkers for its early diagnosis are urgently required. Serum may be a valuable source of biomarkers that may be revealed by proteomic profiling. Herein, complementary serum protein profiling strategies were employed for discovery of biomarkers that could discriminate cases of malignant and benign ovarian cancer.Experimental designIdentically collected and processed serum samples from 22 cases of invasive epithelial ovarian cancer, 45 benign ovarian neoplasms, and 64 healthy volunteers were subjected to immunodepletion and protein equalization coupled to 2D‐DIGE/MS and multidimensional fractionation coupled to SELDI‐TOF profiling with MS/MS for protein identification. Selected candidates were verified by ELISA in samples from malignant (n = 70) and benign (n = 89) cases and combined marker panels tested against serum CA125.ResultsBoth profiling platforms were complementary in identifying biomarker candidates, four of which (A1AT, SLPI, APOA4, VDBP) significantly discriminated malignant from benign cases. However, no combination of markers was as good as CA125 for diagnostic accuracy. SLPI was further tested as an early marker using prediagnosis serum samples. While it rose in cases toward diagnosis, it did not discriminate prediagnosis cases from controls.Conclusions and clinical relevanceThe candidate biomarkers warrant further validation in independent sample sets.

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“…on analytical outcome, particularly for protein and proteomic studies (3,(8)(9)(10)(11). A recent perspective on the subject has been put forward with specific focus on proteome analyses of blood (12).…”

Section: Introductionmentioning

confidence: 99%

Thrombin induces broad spectrum proteolysis in human serum samples

O'Mullan¹,

Craft²,

Yi³

et al. 2009

Clinical Chemistry and Laboratory Medicine

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Background: During clotting, a thrombin cleaves fibrinogen releasing fibrinopeptide A (FPA). FPA is easily identified in serum using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). Using MALDI-TOF MS, we observed multiple, progressively shorter fragments of serum FPA. Following ambient incubation of serum, variations in the content of FPA fragments occur over time. Denaturation of a thrombin by heating the serum sample appears to minimize this variation. These observations suggest that intrinsic proteolytic and peptidolytic activity is elevated in serum and perhaps originates from the coagulation cascade enzymes themselves, especially a thrombin. Methods: Extrinsic addition of a thrombin to a subset (3-30 kDa) of plasma proteins was carried out to induce proteolysis and to examine the resultant peptides to reveal a thrombin susceptible parent proteins. One of these identified proteins, hemopexin, was directly digested by a thrombin and the peptides examined to confirm the observations from the initial plasma protein digestion. Results: Extrinsic addition of a thrombin to a subset (3-30 kDa) of plasma proteins results in wide-spread digestion of proteins unrelated to coagulation, revealing a substrate range encompassing more than fibrinogen. Direct digestion of one of these proteins, hemopexin, by a thrombin confirms these observations. Conclusions: The resulting peptides indicate broad tolerance beyond the consensus R-G cleavage site of fibrinogen; in fact, there appears to be no bias for the amino acid following the R/K residue. These data support our hypothesis that the enzymatic activities inherent to coagulation, or at least to thrombin, contribute to destabilization of the protein and peptide content of serum. Clin Chem Lab Med 2009;47:685-93.

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Influences of Blood Sample Processing on Low–Molecular-Weight Proteome Identified by Surface-Enhanced Laser Desorption/Ionization Mass Spectrometry

Cited by 215 publications

References 46 publications

Clinical proteomics of the cerebrospinal fluid: Towards the discovery of new biomarkers

Clinical proteomics of the cerebrospinal fluid: Towards the discovery of new biomarkers

Discovery of serum biomarkers of ovarian cancer using complementary proteomic profiling strategies

Thrombin induces broad spectrum proteolysis in human serum samples

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