Affinity Capillary Electrophoresis Using a Low-Concentration Additive with the Consideration of Relative Mobilities

Galbusera, Chiara; Thachuk, Mark; Lorenzi, Ersilia De; Chen, David D. Y.

doi:10.1021/ac010875b

Cited by 35 publications

(35 citation statements)

References 25 publications

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“…A bubble cell capillary was utilized in order to increase detection sensitivity due to the relatively poor UV absorption properties of the BS. It is desirable to keep the sample concentrations in mobility shift ACE as low as possible relative to the ligand concentration [27,[40][41][42][43][44] (to be discussed in Section 3.2). To this end a BS concentration of 5610 26 M was found to be detectable and subsequently used as the sample concentration throughout.…”

Section: Ace Proceduresmentioning

confidence: 99%

“…For high affinity interactions other restrictions may apply. In mobility shift ACE experiments, it has been observed that the ligand induced shift in analyte mobility is dependent on the analyte concentration injected [40][41][42][43][44]56]. Consequently, it is generally recommended that a) Stability constants determined by a phase solubility technique (displacement assay) [15].…”

Section: Estimation Of Stability Constantsmentioning

confidence: 99%

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Complexation of tauro‐ and glyco‐conjugated bile salts with three neutral β‐CDs studied by ACE

et al. 2007

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Complexation of the bile salts (BS) taurocholate, tauro-beta-muricholate, taurodeoxycholate, taurochenodeoxycholate, glycocholate, glycodeoxycholate, and glycochenodeoxycholate common in rat, dog, and man with natural beta-CD and the chemically modified beta-CDs 2-hydroxypropyl-beta-CD and 2-O-methyl-beta-CD was studied using mobility shift ACE. The CDs were selected due to their frequent use in preformulation and drug formulation as oral excipients for the solubilization of drug substances with low aqueous solubility. ACE was demonstrated to be a feasible and efficient technique for investigation of the interactions between BS and beta-CDs. All the investigated BS possessed affinity for the three CDs with stability constants ranging from 2x10(3) to 4x10(5) M(-) (1). The requirements and assumptions related to the use of ACE for estimating high affinity stability constants were discussed. The extent and pattern of hydroxylation significantly influenced the affinity of the glyco- and tauro-conjugated BS toward the beta-CDs (chenodeoxycholates >> deoxycholates > cholates) whereas the nature of the beta-CD derivatization and BS conjugation played a minor role only. The results indicate that displacement of drug substances from beta-CD inclusion complexes is likely to occur in the small intestine where BS are present potentially influencing drug bioavailability.

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Section: Ace Proceduresmentioning

confidence: 99%

Section: Estimation Of Stability Constantsmentioning

confidence: 99%

Complexation of tauro‐ and glyco‐conjugated bile salts with three neutral β‐CDs studied by ACE

et al. 2007

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“…On the y-axis are reported the mobility shifts of the isoforms N and I 2 of b 2 -m injected in a capillary filled with (A) 50 mM and (B) 100 mM library compound dissolved in the BGE. Mobility shift is defined as the difference between the effective mobility of b 2 -m peaks in neat buffer and in the presence of the additive [43]; each mobility shift value is the average of two replicates. Inset of (A): electropherograms of b 2 -m injected in the BGE and in the BGE added with 50 mM CREMS-05.…”

Section: Ultrafiltration-cementioning

confidence: 99%

Search of ligands for the amyloidogenic protein β2-microglobulin by capillary electrophoresis and other techniques

et al. 2005

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Beta2-microglobulin (beta2-m) is a small amyloidogenic protein normally present on the surface of most nucleated cells and responsible for dialysis-related amyloidosis, which represents a severe complication of long-term hemodialysis. A therapeutic approach for this amyloidosis could be based on the stabilization of beta2-m through the binding to a small molecule, and consequent inhibition of protein misfolding and amyloid fibril formation. A few compounds have been described to weakly bind beta2-m, including the drug suramin. The lack of a binding site for nonpolypeptidic ligands on the beta2-m structure makes it difficult for both the identification of functional groups responsible for the binding and the search of hits to be optimized. The characterization of the binding properties of suramin for beta2-m by using three different techniques (surface plasmon resonance, affinity CE (ACE), ultrafiltration) is here described and the results obtained are compared. The common features of the chemical structures of the compounds known to bind the protein led us to select 200 sulfonated/suramin-like molecules from a wider chemical library on the basis of similarity rules, so as to possibly single out some interesting hits and to gain more information on the functional groups involved in the binding. The development of screening methods to test the compounds by using ultrafiltration and ACE is described.

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“…When in a CE system at a low concentration of protein as additive a steady state is reached, the additive entering the analyte plug is unaffected by the mobility of the additive. However, when a steady state is not reached, the mobility of the analyte plug may be affected by the amount of additive entering the plug and the time required to reach the steady state [82]. In an ACE study of the kedarcidin-chromophoreapoprotein interaction, the addition of organic solvents was found to reduce the hydrogen bond interaction between the protein and the chromophore [1].…”

Section: Protein-small Molecule Interactionmentioning

confidence: 99%

Recent advances in the study of biomolecular interactions by capillary electrophoresis

Ding

et al. 2004

Electrophoresis

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Recent advances in the study of biomolecular interactions by capillary electrophoresisCapillary electrophoresis (CE) has been abundantly used in the study of molecular interactions owing to such advantages as short analysis time, low sample size requirement, high separation efficiency, and flexible applications. The focus of this paper is to review recent studies and advances (mainly from 1998 to now) in biomolecular interactions using CE. Five CE modes: zone migration CE, affinity CE, frontal analysis (FA), Hummel-Dreyer (HD) and vacancy peak (VP) are cited and compared. Quantitative aspects of the thermodynamics and kinetics of biomolecular interaction are reviewed. Several biomolecular binding systems, including protein-protein (polypeptide), protein-DNA (RNA), protein(polypeptide)-carbohydrate, protein-small molecule, DNAsmall molecule, small molecule-small molecule, have been well characterized by CE. CE is shown to be a powerful tool for the determination of the binding parameters of various bioaffinity interactions.

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Affinity Capillary Electrophoresis Using a Low-Concentration Additive with the Consideration of Relative Mobilities

Cited by 35 publications

References 25 publications

Complexation of tauro‐ and glyco‐conjugated bile salts with three neutral β‐CDs studied by ACE

Complexation of tauro‐ and glyco‐conjugated bile salts with three neutral β‐CDs studied by ACE

Search of ligands for the amyloidogenic protein β2-microglobulin by capillary electrophoresis and other techniques

Recent advances in the study of biomolecular interactions by capillary electrophoresis

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