Automated Multicapillary Electrophoresis for Analysis of Human Serum Proteins

Gay-Bellile, Cécile; Bengoufa, Djaouïda; Houzé, Pascal; Carrer, D. Le; Benlakehal, Mourad; Bousquet, Bernard; Gourmel, Bernard; Bricon, Thierry Le

doi:10.1373/clinchem.2003.017756

Cited by 73 publications

(55 citation statements)

References 30 publications

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“…Human serum and human ascites fluid protein electrophoresis was performed using a Capillarys 2™ CE system (Sebia, Paris, France) that is routinely employed in clinical laboratories [40,41]. Prior to the hydrodynamic injection (4"), 40 µl of serum is automatically diluted 5x in the running buffer (pH 10).…”

Section: Protein Analysis and Capillary Electrophoresismentioning

confidence: 99%

Colloidal stability of nano-sized particles in the peritoneal fluid: Towards optimizing drug delivery systems for intraperitoneal therapy

et al. 2014

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Intraperitoneal (IP) administration of nano-sized delivery vehicles containing small interfering RNA (siRNA) is recently gaining attention as an alternative route for the efficient treatment of peritoneal carcinomatosis. The colloidal stability of nanomatter following IP administration has, however, not been thoroughly investigated yet. Here, enabled by advanced microscopy methods such as Single Particle Tracking (SPT) and Fluorescence Correlation Spectroscopy (FCS), we follow the aggregation and cargo release of nano-scaled systems directly in peritoneal fluids from healthy mice and ascites fluid from a patient diagnosed with peritoneal carcinomatosis. The colloidal stability in the peritoneal fluids was systematically studied in function of the charge (positive or negative) and Poly-Ethylene Glycol (PEG) degree of liposomes and polystyrene nanoparticles, and compared to human serum. Our data demonstrate strong aggregation of cationic and anionic nanoparticles in the peritoneal fluids, while only slight aggregation was observed for the PEGylated ones. PEGylated liposomes, however, lead to a fast and premature release of siRNA cargo in the peritoneal fluids. Based on our observations, we reflect on how to tailor improved delivery systems for IP therapy.

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Section: Protein Analysis and Capillary Electrophoresismentioning

confidence: 99%

Colloidal stability of nano-sized particles in the peritoneal fluid: Towards optimizing drug delivery systems for intraperitoneal therapy

et al. 2014

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“…The capillary electrophoresis of the control serum followed by UV densitometry at 200 nm (Capillarys, Sebia) was considered as a reference [10]. The median relative ratios (experiment was run in 10-fold) between the percentages of the stained electrophoretic fractions (albumin, alpha 1, alpha 2, beta and gamma globulins) and the ones obtained by capillary electrophoresis were calculated to determine the specificity of the Coomassie R-250 and nigrosin staining procedures.…”

Section: Specificity Of the Stainmentioning

confidence: 99%

Development of an affordable dye-stained microalbuminuria screening test

Tugirimana¹,

Delanghe²

2008

Nephrology Dialysis Transplantation

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Methods. Urine was spotted on cellulose acetate strips and stained using different sensitive protein binding dyes (nigrosin, Coomassie Blue R-250, amido black). The colour intensity of the stained spots was quantified using a Kodak Image 450 station.Results. Analytical sensitivity of the Coomassie Blue based method (18 mg/L) was better than that for nigrosin (50 mg/ L) or amido black (100 mg/L) based methods. Within-run coefficient of variation (CV) and between-run CV of the Coomassie blue assay were, respectively, 8.4% and 9.7% (50 mg/L), and 3% and 4.5% (400 mg/L). For nigrosin, these data were, respectively, 8.4 and 9.4 (50 mg/L), and 3.4 and 6.4% (400 mg/L). Coomassie Blue showed a preferential binding selectivity towards albumin. The method was found to be linear between 20 and 600 mg/L. A good corCorrespondence and offprint requests to: Joris R. Delanghe, Department of Clinical Chemistry, De Pintelaan 185, 9000 Gent, Belgium. Tel: +32-9-3322956; Fax: +32-9-332-4985; E-mail: joris.delanghe@ugent.be relation (r 2 = 0.89) was obtained between Coomassie Blue based and immunonephelometric measurements. Immunounreactive albumin (prepared by protease treatment) could be detected by the spot test, which offers an advantage of the method versus immunochemical tests. Ammonium sulphate precipitation could further increase the specificity of the assay by eliminating effects of free light chains. Conclusion. The described method is very simple and extremely cheap, which makes it potentially suited for screening programmes, particularly in third world countries.

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“…First, its high detection limit is not sensitive enough for low burden diseases such as MGRS. The detection limit is 0.3-0.5 g/dL (3-5 g/L) in the γ region and as high as 0.7 g/dL (7 g/L) in α or β region [31]. As a result, SPEP is positive in 87.6% of MM, but only 73.8% of AL and 55.6% of light chain deposition disease (LCDD) [32].…”

Section: Serum Protein Electrophoresismentioning

confidence: 99%

Laboratory testing in monoclonal gammopathy of renal significance (MGRS)

Leung

Barnidge

Hutchison³

2016

Clinical Chemistry and Laboratory Medicine (CCLM)

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Abstract:Recently, monoclonal gammopathy of renal significance (MGRS) reclassified all monoclonal (M) gammopathies that are associated with the development of a kidney disease but do not meet the definition of symptomatic multiple myeloma (MM) or malignant lymphoma. The purpose was to distinguish the M gammopathy as the nephrotoxic agent independent from the clonal mass. The diagnosis of MGRS obviously depends on the detection of the M-protein. More importantly, the success of treatment is correlated with the reduction of the M-protein. Therefore, familiarity with the M-protein tests is a must. Protein electrophoresis performed in serum or urine is inexpensive and rapid due to automation. However, poor sensitivity especially with the urine is an issue particularly with the low-level M gammopathy often encountered with MGRS. Immunofixation adds to the sensitivity and specificity but also the cost. Serum free light chain (sFLC) assays have significantly increased the sensitivity of M-protein detection and is relatively inexpensive. It is important to recognize that there is more than one assay on the market and their results are not interchangeable. In addition, in certain diseases, immunofixation is more sensitive than sFLC. Finally, novel techniques with promising results are adding to the ability to identify M-proteins. Using the time of flight method, the use of mass spectrometry of serum samples has been shown to dramatically increase the sensitivity of M-protein detection. In another technique, oligomeric LCs are identified on urinary exosomes amplifying the specificity for the nephrotoxic M-protein.

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Automated Multicapillary Electrophoresis for Analysis of Human Serum Proteins

Cited by 73 publications

References 30 publications

Colloidal stability of nano-sized particles in the peritoneal fluid: Towards optimizing drug delivery systems for intraperitoneal therapy

Colloidal stability of nano-sized particles in the peritoneal fluid: Towards optimizing drug delivery systems for intraperitoneal therapy

Development of an affordable dye-stained microalbuminuria screening test

Laboratory testing in monoclonal gammopathy of renal significance (MGRS)

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