Enrichment of proteinaceous materials on a strong cation-exchange diol silica restricted access material: protein–protein displacement and interaction effects

Willemsen, Olga; Machtejevas, Egidijus; Unger, Klaus K.

doi:10.1016/j.chroma.2003.10.072

Cited by 27 publications

(13 citation statements)

References 31 publications

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“…As a rule of thumb, the higher amount of sample being injected the more analytes will be detected by the mass spectrometer. As we have demonstrated elsewhere, a possible displacement of analytes can still occur and this will dramatically change the relative amounts of adsorbed species when the column is overloaded [2]. Up to now, we have no proper means to assess a critical loading value.…”

Section: Parameter Adjustment In Sample Clean Up Employing Ram-scx Comentioning

confidence: 89%

“…Thus, both the loadability of the RAM column and its excluding capacity play an essential role in the design of a sample cleanup procedure. Independently, the high abundance ratio of species in the starting sample might lead to displacement and protein -protein complexation, which are difficult to predict and also to control [2].…”

Section: Handling Of Ram-scx Columnsmentioning

confidence: 99%

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Profiling of endogenous peptides by multidimensional liquid chromatography: On‐line automated sample cleanup for biomarker discovery in human urine

Machtejevas¹,

Markó-Varga

Lindberg

et al. 2009

J of Separation Science

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A simple and flexible system, employing a column switching technique, has been designed to allow the analysis of peptides and proteins smaller than 15 kDa by molecular weight in filtered urine samples by performing a direct on-column injection utilising simultaneous sample clean-up and trace enrichment. The positively charged peptides and small proteins in the sample are attracted to the inner, negatively charged pore structure of the RAM-SCX column while the larger proteins and uncharged or negatively charged compounds are excluded. After preconditioning with the biological sample, large amounts of sample can be injected. Several important and adjustable parameters for the proper use of a RAM-SCX column are described and discussed. The main parameters being: i) the column is sensitive to sample overloading, which may result in drastic changes in the adsorption of peptides; ii) adsorption appears to be flow-rate and concentration dependent, as the sample molecules need time to penetrate into the internal pore structure in order to find complimentary orientated adsorption sites; iii) dilution and pH adjustment of sample during the loading process. The biocompatibility and proof-of-principle of this separation platform was demonstrated using human urine samples. Data are presented on repeatability as well as on the reproducibility of different synthesised batches of restricted access material (RAM).

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Section: Parameter Adjustment In Sample Clean Up Employing Ram-scx Comentioning

confidence: 89%

Section: Handling Of Ram-scx Columnsmentioning

confidence: 99%

Profiling of endogenous peptides by multidimensional liquid chromatography: On‐line automated sample cleanup for biomarker discovery in human urine

Machtejevas¹,

Markó-Varga

Lindberg

et al. 2009

J of Separation Science

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“…The handling of extreme differences in abundance is also quite difficult in LC-based approaches and requires experience. [50] Automated sample processing in combination with carefully chosen columns and elution sequences provides a chance to track a large number of sample components. The usefulness of multidimensional LC, including efficient sample clean up procedures, has been shown in specific applications involving various biofluids.…”

Section: Lc In the Postgenomic Eramentioning

confidence: 99%

Liquid Chromatography—Its Development and Key Role in Life Science Applications

et al. 2010

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Liquid chromatographic methods cover the broadest range of applications imaginable today. Nowhere is this more evident and relevant than in the life sciences, where identification of target substances relevant in disease mechanisms is performed down to the femtomole level. On the other hand, purification of therapeutic drugs on a multi-ton scale is performed by process LC. The complexity and abundance range of biological systems in combination with the extreme purity requirements for drug manufacturing are the challenges that can be mastered today by chromatography, after more than a century of research and development. However, significant improvement is still required for a better understanding of the scientific fundamentals of the underlying phenomena and exploiting those for an enhanced quality of live.

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“…Recently several reports concerning the use of cationexchange diol RAM column were reported for analysis of peptides, protein and drugs [10][11][12][13][14]. These reports dealt with the on-line sample clean up of peptides in HPLC-electrospray ionization MS system [10] and the separation of the low-molecular-weight proteinaceous fraction form bio-fluids for protein mapping [11] and size exclusion and enrichment of low molecular mass proteinaceous samples [12].…”

Section: Introductionmentioning

confidence: 99%

Novel methylcellulose-immobilized cation-exchange precolumn for on-line enrichment of cationic drugs in plasma

Yamamoto

Sakaguchi

Kajima

et al. 2004

Journal of Chromatography B

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Enrichment of proteinaceous materials on a strong cation-exchange diol silica restricted access material: protein–protein displacement and interaction effects

Cited by 27 publications

References 31 publications

Profiling of endogenous peptides by multidimensional liquid chromatography: On‐line automated sample cleanup for biomarker discovery in human urine

Profiling of endogenous peptides by multidimensional liquid chromatography: On‐line automated sample cleanup for biomarker discovery in human urine

Liquid Chromatography—Its Development and Key Role in Life Science Applications

Novel methylcellulose-immobilized cation-exchange precolumn for on-line enrichment of cationic drugs in plasma

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