Adaptation of the equipment for high-performance electrophoresis to isoelectric focusing

Hjertén, Stellan; Zhu, Mingde

doi:10.1016/s0021-9673(00)90512-0

Cited by 427 publications

(237 citation statements)

References 9 publications

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“…Capillary isoelectric focusing (CIEF) that is used to separate and concentrate ampholytes such as peptides or proteins based on their isoelectric points (pI), was first reported by Hjerten et al as a highly sensitive analytical method with high resolution (0.02 pI units) [1]. CIEF can be used to analyze small amounts of samples automatically with the help of a commercial apparatus [2], whereas conventional IEF analysis requires a series of tedious manual procedures.…”

Section: Introductionmentioning

confidence: 99%

A Simple and Easy-to-Use Capillary Isoelectric Focusing Technique Using Reagent-Release Hydrogels

et al. 2017

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Reagent-release hydrogels containing an acid or a base solution were prepared and employed as ion suppliers in isoelectric focusing using short capillaries. In conventional isoelectric focusing techniques using microfluidic channels, complicated liquid manipulations are required, which can decrease analytical performance such as the reproducibility of a focusing position. To develop a rapid and sensitive assay based on microfluidic isoelectric focusing, we previously prepared reagent-release capillaries retaining ampholytes by physical adsorption on the inner surface of a short capillary, which were then applied to isoelectric focusing. In this study, reagent-release hydrogels containing an acid or a base solution were developed for isoelectric focusing using short capillaries in place of the acid and base solutions used in conventional microfluidic isoelectric focusing. The prepared reagent-release hydrogels showed some advantages for isoelectric focusing, e.g., high durability for at least five weeks of storage, easy handling as compared to the conventional solutions, and suppression of hydrodynamic flow in capillaries, which often decreases the reproducibility in capillary isoelectric focusing. The migration behavior of ions from the hydrogels into the capillary and the formation of a pH gradient in isoelectric focusing using the reagent-release hydrogels were investigated, which confirmed that the prepared reagent-release hydrogels could form a similar pH gradient to that obtained with conventional capillary isoelectric focusing using the solutions.

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Section: Introductionmentioning

confidence: 99%

A Simple and Easy-to-Use Capillary Isoelectric Focusing Technique Using Reagent-Release Hydrogels

et al. 2017

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“…13 Furthermore, the nano-fabrication schemes need an expensive apparatus and complicated procedures. Thus, it is appropriate for MCE to apply a rapid and simple preconcentration technique including field-amplified stacking (FASS), [16][17][18] sweeping, [19][20][21] transient isotachophoresis, 22,23 isoelectric focusing, 24,25 transient trapping, 26 and so on. Especially, stacking techniques have often been applied to capillary electrophoresis and MCE, since it is easy to use.…”

Section: Introductionmentioning

confidence: 99%

Effect of a Low-conductivity Zone on Field-amplified Sample Stacking in Microchip Micellar Electrokinetic Chromatography

2013

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“…CIEF provides high resolution separation of amphoteric biomolecular species based on differences of their pI in a pH gradient formed by carrier ampholytes, when an electric potential is applied [32]. This high-throughput technique offers greater resolving power and becomes a standard method for the analysis of protein and proteinligand interaction.…”

Section: Introductionmentioning

confidence: 99%

“…This high-throughput technique offers greater resolving power and becomes a standard method for the analysis of protein and proteinligand interaction. The utilization of the whole column detection approach with UV WCID gives this separation technique a unique advantage over the conventional single point detection [32][33][34][35]. For example, in single point detection, several problems are encountered during the sample mobilization process including protein precipitation, long analysis time, and distortion of pH gradient.…”

Section: Introductionmentioning

confidence: 99%

Investigation of interaction between human hemoglobin A₀ and platinum anticancer drugs by capillary isoelectric focusing with whole column imaging detection

Lemma

Mandal

et al. 2008

J of Separation Science

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Original PaperInvestigation of interaction between human hemoglobin A 0 and platinum anticancer drugs by capillary isoelectric focusing with whole column imaging detection CIEF with whole column imaging detection (WCID) was used to investigate the interaction of platinum-based anticancer drugs, cis-platinum(II) diamine dichloride (cisplatin) and [SP-4-2-{1R-trans)]-(1,2-cyclohexanediamine-N,N9)[ethanedioata(2 -)-O,O9]platinum (oxaliplatin), with human hemoglobin A 0 (Hb). This technique facilitates the investigation and characterization of the formation of adducts between drugs and proteins. Cisplatin and oxaliplatin were mixed with the target protein at different concentrations (0:1, 1:1, 1:10, 1:50, and 1:100), and the reaction mixtures were incubated for 0, 0.5, 1, 12, 24, 48, and 72 h at 378C in a water-bath. The focused Hb -drug adduct profiles were imaged by WCID. At higher drug to protein molar ratios (for both oxaliplatin and cisplatin), the results exhibit significant changes in the peak shapes and heights, which may indicate the destabilization of the protein. However, the conformational change was less evident at lower molar ratios. In addition, a major pI shift was observed for the oxaliplatin reaction mixtures (for 1:10, 1:50, and 1:100 ratios). In comparison with previously reported findings obtained by other analytical methods, conclusions were drawn about the validity of CIEF as a simple and convenient method for the investigation of protein -drug interactions. These results may provide useful information for further understanding the activity and toxicity of these chemotherapeutic drugs and improving their clinical performance.

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Adaptation of the equipment for high-performance electrophoresis to isoelectric focusing

Cited by 427 publications

References 9 publications

A Simple and Easy-to-Use Capillary Isoelectric Focusing Technique Using Reagent-Release Hydrogels

A Simple and Easy-to-Use Capillary Isoelectric Focusing Technique Using Reagent-Release Hydrogels

Effect of a Low-conductivity Zone on Field-amplified Sample Stacking in Microchip Micellar Electrokinetic Chromatography

Investigation of interaction between human hemoglobin A₀ and platinum anticancer drugs by capillary isoelectric focusing with whole column imaging detection

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Adaptation of the equipment for high-performance electrophoresis to isoelectric focusing

Cited by 427 publications

References 9 publications

A Simple and Easy-to-Use Capillary Isoelectric Focusing Technique Using Reagent-Release Hydrogels

A Simple and Easy-to-Use Capillary Isoelectric Focusing Technique Using Reagent-Release Hydrogels

Effect of a Low-conductivity Zone on Field-amplified Sample Stacking in Microchip Micellar Electrokinetic Chromatography

Investigation of interaction between human hemoglobin A0 and platinum anticancer drugs by capillary isoelectric focusing with whole column imaging detection

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Investigation of interaction between human hemoglobin A₀ and platinum anticancer drugs by capillary isoelectric focusing with whole column imaging detection