Protein reducing agents dithiothreitol and tris(2-carboxyethyl)phosphine anodic oxidation

Santarino, Inês B.; Oliveira, Severino Carlos B.; Oliveira-Brett, Ana Maria

doi:10.1016/j.elecom.2012.06.027

Cited by 31 publications

(16 citation statements)

References 9 publications

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“…DP voltammograms of 50 mM DTT in phosphate buffer showed only one oxidation peak 1 a , at E pa = +1.15 V (Figure 2 A) due to the oxidation of the sulfhydryl group with lowest pK a [19]. DP voltammograms of 50 mM TCEP in phosphate buffer showed also only one oxidation peak 1 a , at E pa = + 0.94 V (Figure 2 B) due to the oxidation of the phosphine group in the molecule [19]. DP voltammograms were also recorded for 1 mM SDS solution in phosphate buffer pH 7.0, but no oxidation peak was detected.…”

Section: Special Issuementioning

confidence: 98%

“…TCEP and DTT have been applied to protein samples with the aim to cleave the disulfide bond between cysteine residues [19], and TCEP is considered to have at pH < 8 a higher reducing power in comparison with DTT [22].…”

Section: Special Issuementioning

confidence: 99%

“…The electrochemical oxidation mechanism of the reducing agents DTT and TCEP was investigated using cyclic and differential pulse voltammetry over a wide pH range on a glassy carbon electrode [19]. The denaturing agent SDS is not electroactive.…”

Section: Oxidation Of Denatured Rtxmentioning

confidence: 99%

“…Due to protein structural complexity, the use of the denaturing SDS, a detergent that dissolves hydrophobic proteins, and the reducing agents DTT or TCEP, to disrupt disulfide bonds, are essential for qualitative and quantitative analysis in proteomic mechanism investigation [19,20].…”

Section: Oxidation Of Denatured Rtxmentioning

confidence: 99%

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Direct Electrochemistry of Native and Denatured Anticancer Antibody Rituximab at a Glassy Carbon Electrode

2013

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Rituximab (RTX) is a human/murine chimeric monoclonal antibody (mAb) that specifically targets the transmembrane protein CD20 of B-cells. The oxidation mechanism of native and denatured RTX was investigated on glassy carbon electrode. The denaturing agent sodium dodecyl sulfate and the redutancts tris(2-carboxyethyl)phosphine and dithiothreitol were used. Significant differences were observed for native and denatured RTX oxidation due to morphological changes and unfolding of the RTX native structure. Native RTX presented only one oxidation peak of tyrosine and tryptophan residues, whereas in denatured RTX were detected three peaks corresponding to the oxidation of tyrosine, tryptophan and histidine residues.

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Section: Special Issuementioning

confidence: 98%

Section: Special Issuementioning

confidence: 99%

Section: Oxidation Of Denatured Rtxmentioning

confidence: 99%

Section: Oxidation Of Denatured Rtxmentioning

confidence: 99%

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Direct Electrochemistry of Native and Denatured Anticancer Antibody Rituximab at a Glassy Carbon Electrode

2013

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“…Electrochemical techniques have been applied to help clarifying such drug mechanism of action. Moreover, due to their high sensitivity, voltammetric methods have been successfully used for detection and determination of various pharmaceutical and hazardous compounds [4][5][6][7][8][9].…”

Section: Introductionmentioning

confidence: 99%

Electrochemical Oxidation Mechanism of Ethidium Bromide at a Glassy Carbon Electrode

Oliveira

Nascimento

2013

Electroanalysis

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The electrochemical behavior of ethidium bromide (EtBr) on a glassy carbon electrode (GCE) was studied by cyclic, square wave and differential pulse voltammetry over a wide pH interval. The results revealed that the oxidation mechanism of EtBr is an irreversible and adsorption-controlled electrode process that occurs in two consecutives steps. The first step is pH-dependent and occurs at the amino group in the C8 position with the formation of ortho-and para-quinone derivatives, while the second step is pH-independent and occurs at the amino group in the C3 position. A square wave method for quantitative determination of EtBr is also proposed.

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LC–MS/MS determination of N‐acetyl‐l‐cysteine in chicken plasma

Roydeva,

Milanova

2024

Biomedical Chromatography

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N‐acetyl‐l‐cysteine (NAC) shows beneficial effects in cases of aflatoxicosis and heat stress in poultry but little is known about its pharmacokinetics in chickens. Therefore, the study aimed to develop and validate a sensitive LC–MS/MS analytical method for quantitative analysis of NAC in chicken plasma. A split calibration curve approach was used for determination of NAC in chicken plasma. Standard curves for low (0.05–2.5 μg/ml) and high (2.5–100 μg/ml) ranges of concentrations were prepared. The standard curves for low (r2 = 0.9987) and high (r2 = 0.9899) concentrations were linear within the tested range. The limits of detection (LOD) and of quantification (LOQ) for the standard at low concentrations were 0.093 and 0.28 μg/ml, respectively. The accuracy was from 97.35 to 101.33%. The values of LOD and LOQ for the standard at high concentrations were 0.76 and 2.30 μg/ml, respectively. The accuracy was between 99.77 and 112.14%. The intra‐ and inter‐day precisions for all concentrations from both standards did not exceed 8.57% and 10.69%, respectively. The recovery for all concentrations was between 92.45 and 105.52%. The validated method for determination of NAC in chicken plasma can be applied in future pharmacokinetic studies in chickens without dilution of samples and their repeated analysis.

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Protein reducing agents dithiothreitol and tris(2-carboxyethyl)phosphine anodic oxidation

Cited by 31 publications

References 9 publications

Direct Electrochemistry of Native and Denatured Anticancer Antibody Rituximab at a Glassy Carbon Electrode

Direct Electrochemistry of Native and Denatured Anticancer Antibody Rituximab at a Glassy Carbon Electrode

Electrochemical Oxidation Mechanism of Ethidium Bromide at a Glassy Carbon Electrode

LC–MS/MS determination of N‐acetyl‐l‐cysteine in chicken plasma

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