Analytical techniques for cell fractions

Anderson, Norman G.; Tollaksen, Sandra L.; Hahn, H.; Giere, Frederic A.; Edwards, JJ

doi:10.1016/0003-2697(79)90184-2

Cited by 53 publications

(6 citation statements)

References 17 publications

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“…Earlier studies demonstrated an increase in the number of identified proteins in urine through the use of antibody-based depletion of abundant proteins [14] and deglycosylation of urinary proteins [20]. In this study, in addition to the depletion and the deglycosylation, we utilized ultrafiltration of the depleted urine samples, and fractional elution of the peptides from SPE adsorbent (Fig.…”

Section: Discussionmentioning

confidence: 99%

Exploratory study of proteins in urine of patients with histoplasma antigenuria

Kushnir

Crockett

Cloud

et al. 2012

Journal of Chromatography B

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Section: Discussionmentioning

confidence: 99%

Exploratory study of proteins in urine of patients with histoplasma antigenuria

Kushnir

Crockett

Cloud

et al. 2012

Journal of Chromatography B

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“…Therefore the proteins of urine must be separated from the majority of the nonprotein components prior to 2-DE. Various preparatory methods have been described for desalting and concentration, including dialysis and gel chromatography [23-251, gel chromatography and centrifugation [24], dialysis against PEG [26] and an acidified acetone extraction with subsequent size exclusion chromatography [27]. Tracy and coworkers recommend the combination of size exclusion chromatography under high pressure (SE-HPLC) followed by lyophilization of the protein eluate [2].…”

Section: Discussionmentioning

confidence: 99%

Electrophoretic, chromatographic and immunological studies of human urinary proteins

Büeler¹,

Wiederkehr²,

Vonderschmitt³

1995

Electrophoresis

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Urinary proteins from both sexes were analyzed by high resolution two-dimensional gel electrophoresis (2-DE). For well reproducible 2-DE patterns, the samples were concentrated and desalted in one step by vacuum dialysis. A reference map for urine proteins was established by the analysis of urine from 10 healthy persons. Proteins in urine that share immunogenicity with serum proteins were identified by use of antibody to whole human-serum protein in an affinity-column fractionation of urine and differential analysis of the adsorbed (serum component) and unadsorbed (non-serum component) fractions. For identification of individual proteins, coelectrophoresis, immunoblotting and affinity chromatography with corresponding antibodies were used. Proteins identified in the map, besides known serum proteins, included: the subunit of Tamm-Horsefall protein, the secretory component of IgA, constant breakdown products of alpha 1-antitrypsin and retinol-binding protein, the five isoforms of the beta chain of human chorionic gonadotropin and the subunit of prostatic acid phosphatase. In addition, we could demonstrate three proteins which are markedly pronounced in female urine, especially pregnant women. To get more information about the native properties of various urinary proteins, they were separated into four main peaks according to their sizes using fast protein liquid chromatography equipment. Possible interpolypeptide disulfide bonds were studied using a nonreducing 2-DE system. 2-DE in combination with other methods seems to be a valuable tool for the characterization of urinary proteins in defined renal or extra-renal diseases. An example is given by analyzing the immune complexes from seven patients with a urinary tract infection.

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“…High efficient CE methods have recently been taken up by clinical laboratory to replace gel electrophoresis for routine monitoring with related methodologies validated. However, time-consuming sample pretreatment to remove non-protein substances in urine samples are still required to achieve a sufficiently high detection sensitivity to determine minor urinary proteins such as b2-MG [2,10,11]. Thus, coupling CE with a microchip capable of online sample cleanup and preconcentration prior to CZE separation of urinary protein in an embedded capillary is investigated.…”

Section: Microchip-ce Device For Cze Separation Of Urinary Proteinsmentioning

confidence: 99%

“…To solve the interfering problems due to salts and low molecular weight metabolic wastes, both chemical and physical processes have been used for pretreatment of urine samples, including dialysis [10], ultrafiltration [11] and precipitation [12]. These methods can effectively remove non-protein materials.…”

Section: Introductionmentioning

confidence: 99%

2‐D t‐ITP/CZE determination of clinical urinary proteins using a microfluidic‐chip capillary electrophoresis device

Yeung

Fung

2011

Electrophoresis

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To replace the time-consuming sample pretreatment procedure, a microfluidic chip-CE device incorporating on-chip sample desalting/preconcentration with transient isotachophoresis (ITP)/capillary zone electrophoresis (CZE) was fabricated to perform sequential on-chip sample pretreatment and CE determination of four urinary proteins in clinical samples. On-chip sample desalting, clean-up and analyte preconcentration enable removing interfering sample matrix prior to transferring analytes to separation capillary for transient ITP/CZE determination. Four important urinary proteins transferrin, β2-microglobulin, human serum albumin (HSA) and immunoglobulin G (IgG), investigated were shown to achieve quantitation limits sufficiently high to meet medical requirements, sensitivity enhancement up to 40-fold and detection limits down to 0.3, 0.05, 0.6, 0.5 mg/L, respectively. Compared to the stacking effect, the use of a large sample size was found to be the major factor for sensitivity enhancement. The method reliability is established by close to 100% recoveries and statistical agreement of results from the method developed with currently used clinical radio-immunoassay method for all four proteins investigated. Moreover, an assay time of less than 10 min is needed in the method developed as compared to 7 h for the radio-immunoassay method.

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Analytical techniques for cell fractions

Cited by 53 publications

References 17 publications

Exploratory study of proteins in urine of patients with histoplasma antigenuria

Exploratory study of proteins in urine of patients with histoplasma antigenuria

Electrophoretic, chromatographic and immunological studies of human urinary proteins

2‐D t‐ITP/CZE determination of clinical urinary proteins using a microfluidic‐chip capillary electrophoresis device

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