Development of optimized mobile phases for protein separation by high performance thin layer chromatography

Biller, Julia; Morschheuser, Lena; Riedner, Maria; Rohn, Sascha

doi:10.1016/j.chroma.2015.08.048

Cited by 18 publications

(13 citation statements)

References 31 publications

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“…4B,C, lane 1), only lysozyme shows a separation (R f 0.29). As reported by Biller et al ., thin-layer chromatographic separation of ovalbumin cannot be completely accomplished under these conditions4. However, a clean-up effect of the sample was achieved resulting in a distinct lysozyme band.…”

Section: Resultsmentioning

confidence: 99%

“…To overcome some of the problems while using LC-MS or electrophoresis, high-performance thin-layer chromatography (HPTLC) represents an interesting complementary technique1234. In contrast to the dominantly used electrophoretic approaches, more degrees of freedom with regard to separation and detection are available.…”

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confidence: 99%

“…The analysis of non-denatured proteins, especially proteoforms, is still challenging due to their multitude of structural and physicochemical characteristics. To overcome some of the problems while using LC-MS or electrophoresis, high-performance thin-layer chromatography (HPTLC) represents an interesting complementary technique 1 2 3 4 . In contrast to the dominantly used electrophoretic approaches, more degrees of freedom with regard to separation and detection are available.…”

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confidence: 99%

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HPTLC-aptastaining – Innovative protein detection system for high-performance thin-layer chromatography

Morschheuser¹,

Wessels²,

Pille³

et al. 2016

Sci Rep

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Protein analysis using high-performance thin-layer chromatography (HPTLC) is not commonly used but can complement traditional electrophoretic and mass spectrometric approaches in a unique way. Due to various detection protocols and possibilities for hyphenation, HPTLC protein analysis is a promising alternative for e.g., investigating posttranslational modifications. This study exemplarily focused on the investigation of lysozyme, an enzyme which is occurring in eggs and technologically added to foods and beverages such as wine. The detection of lysozyme is mandatory, as it might trigger allergenic reactions in sensitive individuals. To underline the advantages of HPTLC in protein analysis, the development of innovative, highly specific staining protocols leads to improved sensitivity for protein detection on HPTLC plates in comparison to universal protein derivatization reagents. This study aimed at developing a detection methodology for HPTLC separated proteins using aptamers. Due to their affinity and specificity towards a wide range of targets, an aptamer based staining procedure on HPTLC (HPTLC-aptastaining) will enable manifold analytical possibilities. Besides the proof of its applicability for the very first time, (i) aptamer-based staining of proteins is applicable on different stationary phase materials and (ii) furthermore, it can be used as an approach for a semi-quantitative estimation of protein concentrations.

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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HPTLC-aptastaining – Innovative protein detection system for high-performance thin-layer chromatography

Morschheuser¹,

Wessels²,

Pille³

et al. 2016

Sci Rep

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“…(6) and (9)]. After having examined the influence of solvents from different Snyder groups, the relationships between the mobile phase properties and the retention behavior of ziprasidone and its impurities were investigated by correlating the obtained migration distances as dependent variables with the calculated mobile phase polarity or solubility values as independent variables.…”

Section: Correlation Between Retention Behavior and Selected Mobile Pmentioning

confidence: 99%

“…Literature survey has revealed numerous examples of application of the Snyder solvent classification system at an initial step of the TLC method optimization. [4][5][6] In addition to polarity, the solubility parameter as an important criterion in terms of selection of a suitable mixture of solvents could also be considered. Hansen presented the concept of total solubility as a sum of the dispersive (δ d ), polar (δ p ) (Keesom and Debye), and hydrogen bonding (δ h ) (donor and acceptor) contributions.…”

Section: Introductionmentioning

confidence: 99%

Influence of selected mobile phase properties on the TLC retention behavior of ziprasidone and its impurities

Obradović

Oljačić

Nikolić

et al. 2017

Journal of Liquid Chromatography & Related Technologies

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Influence of nine different solvents, either alone or in a mixture, on the retention behavior of ziprasidone and its five impurities were examined by normal-phase thin-layer chromatography. Migration distances of the examined compounds obtained under the examined chromatographic conditions were correlated with calculated mobile phase properties, such as Snyder polarity and Hansen solubility. Linear or second-order polynomial relationships with high correlation coefficients were established between investigated variables. The obtained mathematical functions and statistical results indicated that selected mobile phase properties can be used for the prediction of the retention behavior of ziprasidone and its five impurities. GRAPHICAL ABSTRACT

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Massenspektrometrische Identifizierung von Peptid‐ und Proteinbanden aus Molkenprotein‐angereichertem Käse nach dünnschichtchromatographischer Trennung

Treblin¹,

Class²,

Kuhnen³

et al. 2020

Lebensmittelchemie

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Development of optimized mobile phases for protein separation by high performance thin layer chromatography

Cited by 18 publications

References 31 publications

HPTLC-aptastaining – Innovative protein detection system for high-performance thin-layer chromatography

HPTLC-aptastaining – Innovative protein detection system for high-performance thin-layer chromatography

Influence of selected mobile phase properties on the TLC retention behavior of ziprasidone and its impurities

Massenspektrometrische Identifizierung von Peptid‐ und Proteinbanden aus Molkenprotein‐angereichertem Käse nach dünnschichtchromatographischer Trennung

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