A simple affinity spin tube filter method for removing high‐abundant common proteins or enriching low‐abundant biomarkers for serum proteomic analysis

Wang, Young Y.; Cheng, Paul; Chan, Daniel W.

doi:10.1002/pmic.200390036

Cited by 110 publications

(66 citation statements)

References 15 publications

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“…Intact as well as partially degraded proteins or protein fragments circulate in the blood. Furthermore, genetic polymorphisms as well as numerous post-translationally modified forms of proteins are present, [17][18][19][20]. Removal of fibrinogen (Factor I) is easily obtained by clotting.…”

Section: Overviewmentioning

confidence: 99%

Recent advances in blood‐related proteomics

et al. 2005

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Service régional vaudois de transfusion sanguine, Lausanne, SwitzerlandBlood is divided in two compartments, namely, plasma and cells. The latter contain red blood cells, leukocytes, and platelets. From a descriptive medical discipline, hematology has evolved towards a pioneering discipline where molecular biology has permitted the development of prognostic and diagnostic indicators for disease. The recent advance in MS and protein separation now allows similar progress in the analysis of proteins. Proteomics offers great promise for the study of proteins in plasma/serum, indeed a number of proteomics databases for plasma/ serum have been established. This is a very complex body fluid containing lipids, carbohydrates, amino acids, vitamins, nucleic acids, hormones, and proteins. About 1500 different proteins have recently been identified, and a number of potential new markers of diseases have been characterized. Here, examples of the enormous promise of plasma/serum proteomic analysis for diagnostic/prognostic markers and information on disease mechanism are given. Within the blood are also a large number of different blood cell types that potentially hold similar information. Proteomics of red blood cells, until now, has not improved our knowledge of these cells, in contrast to the major progresses achieved while studying platelets and leukocytes. In the future, proteomics will change several aspects of hematology.

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Section: Overviewmentioning

confidence: 99%

Recent advances in blood‐related proteomics

et al. 2005

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“…Many commercial affinity albumin removal kits have been designed specifically for use with serum/CSF and for a minimum of unspecific binding. Among the albumin depletion columns, Cibacron Blue-Sepharose (73) media or monoclonal antibodies (74), (75), (76) are the most common affinity materials. The protein concentration is much higher in serum than in CSF and reducing the volume of CSF might be necessary before the affinity removal of albumin (77).…”

Section: Prefractionation Techniquesmentioning

confidence: 99%

Prefractionation of Cerebrospinal Fluid to Enhance Glycoprotein Concentration Prior to Structural Determination with FT−ICR Mass Spectrometry

2005

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När vi leva, låtom oss leva. While we live, let us live. "Let us enjoy life." Dum vivimus vivamus.3 Mass spectrometry for comparative proteomics of degenerative and regenerative processes in the brain ABSTRACT Biological processes involve changes at the protein level which can be detected and quantified. Proteomics aims to determine protein changes from a normal state, for instance to measure the degree of recovery in a biological system or the state of disease progression. Mass spectrometry is the most important tool in proteomics for the identification of proteins and determination of post-translational modifications such as glycosylation. Glycoproteins were found to be altered in patients with Alzheimer's disease (AD), which is the most common form of dementia. Changes in glycosylation levels were quantified with a glycoprotein-specific stain after gel separation. Glycan structures were determined with mass spectrometry in this thesis. Protein quantification with mass spectrometric methods is based on stable isotope labeling of proteins and labeled cell cultures can be used as internal standards for tissue proteomics. Quantitative proteomics was applied to assess protein expression levels with mass spectrometry in the murine brain after specific neurosurgery to study regenerative processes.In order to compare glycosylated proteins in cerebrospinal fluid (CSF) from individual AD patients with healthy control individuals, glycoproteomic methods were developed. To enhance the concentration of glycoproteins prior to gel separation, affinity chromatography of CSF was the most suitable prefractionation method for removing the most abundant CSF protein albumin. CSF proteins were separated with narrow pH-range two-dimensional gel electrophoresis followed by multiple staining for quantification of glycoprotein isoforms. Structural analysis of glycopeptides was performed with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS), which provided very high mass accuracy and facilitated site-specific determination. A decreased glycosylation level for a protein localized in senile plaques, α 1 -antitrypsin, was found in AD patients. No specific glycoform of the studied proteins could be assigned to AD, emphasizing that further studies should include a larger subject group and cover proteins in various pH intervals. Knowledge of the respective glycoprotein structures in relation to clinical disease parameters may assist in the elucidation of the pathogenesis.In order to study proteins involved in the response of astrocytes and regenerative processes after neurotrauma, a quantitative mass spectrometric method was developed. Astrocytes, which are the most abundant cells in the central nervous system, react to neurotrauma by becoming reactive (reactive gliosis). Mice lacking the intermediate filament proteins GFAP and vimentin (GFAP -/-Vim -/-) show attenuated reactive gliosis and enhanced regeneration after neurotrauma. Comparative proteomic analysis showed upregulation of the adapter protein 14-3-3 in wildt...

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“…Cibacron Blue Dye and protein A/G columns have been used to effectively remove serum albumin and immunoglobulins from serum, allowing detection of proteins present at very low concentrations. Nowadays, the most common technique is the immunodepletion, which has been extensively used for the specific removal of the most abundant proteins, based on the action of specific antibodies (Zolotarjova N. et al, 2005 andWang YY et al, 2003). Among these, Multiple Affinity Removal Columns (MARC) is the most effective method because it simultaneously removes multiple abundant proteins (Bjorhall K, et al, 2005 andSeam N et al, 2007).…”

Section: As Plasma Proteomementioning

confidence: 99%