Pre-column derivatization of proteins to enhance detection sensitivity for sodium dodecyl sulfate non-gel sieving capillary electrophoresis

Gump, Edwin; Monnig, Curtis A.

doi:10.1016/0021-9673(95)00591-a

Cited by 33 publications

(14 citation statements)

References 32 publications

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“…Precolumn derivatization of proteins with fluorescence labels enhances detection sensitivity for SDS-CGE of proteins. Gump and Monning [27] used fluorescamine, naphthalene-2,3-dicarboxyaldehyde and o-phthaldialdehyde to increase detection sensitivity in both UV absorption and fluorescence detection modes. They found an enhancement factor of 22 using UV detection at 280.…”

Section: Detectionmentioning

confidence: 99%

Capillary sodium dodecyl sulfate‐gel electrophoresis of proteins

Guttman¹

1996

Electrophoresis

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Section: Detectionmentioning

confidence: 99%

Capillary sodium dodecyl sulfate‐gel electrophoresis of proteins

Guttman¹

1996

Electrophoresis

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“…[21,22] This probe reacted with primary amines, giving compounds fluorescent at 410 nm (see Fig. 2, reaction 3).…”

Section: Derivatization Conditions and Sample Preparationmentioning

confidence: 99%

Analysis of Amphetamine Derivatives in Plasma Using Capillary Zone Electrophoresis Coupled with Laser-Induced Fluorescence after Derivatization on Solid-Phase Extraction Support

Gremaud¹,

Schappler²,

Veuthey³

et al. 2008

Chimia

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Capillary electrophoresis (CE) is a very powerful analytical technique, consuming very small quantities of samples and solvents. Generally, a UV detector is coupled with CE but for the analysis of drugs in biological fluids, it is necessary to use a more sensitive and selective detector such as laser induced fluorescence (LIF). This detector was used to analyze various amphetamine derivatives which do not present native fluorescence and therefore a derivatization tag was employed to produce a fluorescent adduct. Fluorescamine, a fluorogene agent, was selected because it reacts with primary amines of amphetamines to give a fluorescent compound negatively charged, which can be analyzed by CZE-LIF. Ta king into account the presence of many interfering compounds in biological fluids, analysis of these compounds requires a sample preparation prior to the analysis. Solid phase extraction (SPE) was selected and the derivatization step included. Analysis of independent plasma samples (n = 6) exhibited a relative standard deviation (RSD) of 8%. A relative recovery of 99% was observed for amphetamine, used as a model compound.

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“…Gump and Monnig [56] have reported using OPA, fluorescamine, and NDA to derivatize proteins with fluorescence detection to enhance sensitivity. Hunt and Nashabeh [53] have reported a method where proteins are derivatized using a fluorophore and the resulting derivatives are detected using laser-induced fluorescence resulting in sensitivity similar to silver staining.…”

Section: Sensitivitymentioning

confidence: 99%

Optimization, Validation, and Use of Capillary Gel Electrophoresis for Quality Control Testing of Synagis®, a Monoclonal Antibody

Schenerman¹,

Bowen²

2001

CE in Biotechnology: Practical Applications for Protein and Peptide Analyses

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Capillary electrophoresis methods offer many advantages over their conventional electrophoresis counterparts including: greater precision, higher throughput, on-line detection and less waste. Capillary gel electrophoresis (CGE), using replaceable sieving polymers, is Widely used as an alternative to SDS-PAGE and densitometry CGE offers greater automation capabilities than SDS-PAGE and can be operated using conventional chromatography system software. This software for handling and analyzing data makes the validation of CGE methods look quite similar to the validation of an HPLC method. The same parameters can be examined, including: precision, accuracy, linearity, selectivity, range, robustness, and system suitability. There are, however, certain key elements that are critical to a successful CGE method validation. The CGE quantification method must be shown to be equivalent to SDS-PAGE with densitometry and should be presented such that the densitometry plots look like the electropherograms. In addition, the sensitivity of the methods must be compared and the CGE method must have comparable or better sensitivity to SDS-PAGE with Coomassie staining. The validation must also be performed using materials that might be tested during the course of a typical process (in-process samples) in their particular matrices. The other vital element is assuring that acceptance criteria for the validation have been set to reasonable ranges. These ranges must be determined in pilot studies that are executed prior to the validation studies. Under most CGE conditions, the acceptance criteria will be quite similar to those seen in HPLC validations. Overall, CGE methods can be validated to the same level of assurance as HPLC methods. The CGE method described here has been approved by the FDA and is used routinely for testing of Synagis®.

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Pre-column derivatization of proteins to enhance detection sensitivity for sodium dodecyl sulfate non-gel sieving capillary electrophoresis

Cited by 33 publications

References 32 publications

Capillary sodium dodecyl sulfate‐gel electrophoresis of proteins

Capillary sodium dodecyl sulfate‐gel electrophoresis of proteins

Analysis of Amphetamine Derivatives in Plasma Using Capillary Zone Electrophoresis Coupled with Laser-Induced Fluorescence after Derivatization on Solid-Phase Extraction Support

Optimization, Validation, and Use of Capillary Gel Electrophoresis for Quality Control Testing of Synagis®, a Monoclonal Antibody

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