Peptide Analysis by Rapid, Orthogonal Technologies with High Separation Selectivities and Sensitivities

Hearn, Milton Thomas William

doi:10.1006/biol.2001.0309

Cited by 46 publications

(27 citation statements)

References 66 publications

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“…Furthermore, the resolution was in the range of 1.01-1.51 in acid BGEs and the samples were not baseline-separated in BGE II and BGE III. The analogs with a longer peptide chain, decapeptides, YDPAP 6 and YDPAP 5 DP, and pentapeptides, YDPAP and YDPADP, were completely baseline-separated in all acid BGEs. On the other From CZE analyses of IOPs their effective electrophoretic mobilities were determined.…”

Section: Separation Of Peptide Mixtures and Determination Of Peptide mentioning

confidence: 97%

“…Better values of correlation coefficients were obtained but from the regression analysis it was evident, that di-and tripeptide did not fit well in this linear part of the graphs. Due to nonlinearity of the correlations between effective mobility and charge-tosize ratio of these two short peptides and with respect to their low potential to form more organized secondary structures, the set of IOPs was further reduced to the series of seven peptides with linearly decreasing number of proline residues at the C-terminal of the peptide chain, from decapeptide, YDPAP 6 , to tetrapeptide, YDPA, with constant decrement of the relative molecular mass, 97.1. The graphs of semiempirical models of correlations of electrophoretic mobility and effective charge and size parameters applied to this series of seven linear IOPs are depicted in Fig.…”

Section: Testing Semiempirical Modelsmentioning

confidence: 99%

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Separation and investigation of structure‐mobility relationships of insect oostatic peptides by capillary zone electrophoresis

et al. 2004

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Capillary zone electrophoresis (CZE) has been applied to qualitative analysis, separation, and physicochemical characterization of synthetic insect oostatic peptides (IOPs) and their derivatives and fragments. Series of homologous IOPs were separated in three acidic background electrolytes (BGEs; pH 2.25, 2.30, 2.40) and an alkaline BGE (pH 8.1). Best separation was achieved in acid BGE composed of 100 mM H3PO4, 50 mM Tris, pH 2.25. The effective electrophoretic mobilities, mu(ep), of all IOPs in four BGEs were determined and several semiempirical models correlating effective mobility with charge-to-size ratio (mu(ep) versus q/Mr k) were tested to describe the migration behavior of IOP in CZE. None of models was found to be unambiguously applicable for the whole set of 20 IOPs differing in size (dipeptide - decapeptide) and charge (-2 to +0.77 elementary charges). However, a high coefficient of correlation, 0.9993, was found for the subset of homologous series of IOPs with decreasing number of proline residues at C-terminus, H-Tyr-Asp-Pro-Ala-Prox-OH, x = 6 - 0, for the dependence of mu(ep) on q/Mr k with k = 0.5 for IOPs as anions in alkaline BGE and with k = 2/3 for IOPs as cations in optimized acidic Tris-phosphate BGE. From these dependences the probable structure of IOPs in solution could be predicted.

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Section: Separation Of Peptide Mixtures and Determination Of Peptide mentioning

confidence: 97%

Section: Testing Semiempirical Modelsmentioning

confidence: 99%

Separation and investigation of structure‐mobility relationships of insect oostatic peptides by capillary zone electrophoresis

et al. 2004

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“…Moreover, sample pretreatment (such as solid-phase extraction) is often necessary prior to analysis. CE offers several advantages over chromatographic techniques for the analysis of biological samples [6][7][8]. The main advantages of CE over chromatographic methods are higher separation efficiency, higher selectivity, the smaller sample volume required, and minimal sample processing before injection.…”

Section: Introductionmentioning

confidence: 99%

Separation and determination of carnosine-related peptides using capillary electrophoresis with laser-induced fluorescence detection

et al. 2005

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A capillary electrophoresis (CE) method with laser-induced fluorescence (LIF) detection was developed for the separation and detection of carnosine-related peptides (carnosine, anserine, and homocarnosine). A sensitive and fluorogenic regent, 3-(4-carboxybenzoyl) quinoline-2-carboxaldehyde (CBQCA) was selected as a precapillary labeling reagent for imidazole dipeptides to form isoindole derivatives. The optimized molar ratio between CBQCA and peptide was found to be 75:1, and 50 mmol/L borate buffer (pH 9.2) was used for the derivatization in order to achieve good efficiency. Three imidazole dipeptides were baseline-separated within 20 min by using 112 mmol/L sodium borate (pH 10.4-10.8) as running buffer. Concentration detection limits (signal-to-noise ratios) for carnosine, anserine, and homocarnosine were 4.73, 4.37, and 3.94 nmol/L, respectively. This method has been applied to the analysis of human cerebrospinal fluid (CSF) and meat dry powder of pig and sheep. Recoveries were in the range of 82.9-104.8% for homocarnosine in CSF. For carnosine and anserine, the recoveries are 98.3% and 80.2% in meat dry powder of pig and 111.2% and 112.8% in meat dry powder of sheep, respectively.

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“…In fact, because the basis of the separation is not at all related to hydrophobicity, CE is, in many ways, ideal for separating peptide mixtures that may consist of at least some large (> 20 aa) peptides [20]. Acidic conditions are best for peptides.…”

Section: Separation Of Neuropeptidesmentioning

confidence: 99%

Techniques for neuropeptide determination

Sandberg

Weber

2003

TrAC Trends in Analytical Chemistry

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Because immunoassay responds to epitopes, and many molecules share the same peptide epitope, it is very difficult to obtain an accurate understanding of peptides, their creation and hydrolysis, in biological systems. Separate-and-detect approaches have merit in that the many active peptides and inactive fragments of a particular system can be separately determined. This review discusses the separation, by chromatography and capillary electrophoresis, and detection, by absorbance, fluorescence, electrochemistry, and immunoassay techniques. When separation pre-concentration is accompanied by laser-induced fluorescence or biuret-based electrochemical detection, nM-pM detection limits are obtained.

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Peptide Analysis by Rapid, Orthogonal Technologies with High Separation Selectivities and Sensitivities

Cited by 46 publications

References 66 publications

Separation and investigation of structure‐mobility relationships of insect oostatic peptides by capillary zone electrophoresis

Separation and investigation of structure‐mobility relationships of insect oostatic peptides by capillary zone electrophoresis

Separation and determination of carnosine-related peptides using capillary electrophoresis with laser-induced fluorescence detection

Techniques for neuropeptide determination

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