Comparison of five methods for the study of drug–protein binding in affinity capillary electrophoresis

Busch, Markus; Carels, L.B.; Boelens, H.F.M.; Kraak, J.C.; Poppe, H.

doi:10.1016/s0021-9673(97)00369-5

Cited by 227 publications

(236 citation statements)

References 26 publications

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“…The method can be applied over a very wide pH range (pH [1][2][3][4][5][6][7][8][9][10][11][12][13][14], and be operated with aqueous media. Therefore, CE is suited to study almost any biomolecular interaction system.…”

Section: Discussionmentioning

confidence: 99%

“…The distinction between strong and weak affinity interactions in CE studies does rest not only on a fixed value of the binding constant but also on the experimental setup, primarily on the separation time and buffer conditions [10]. Five CE modes including zone migration CE (CZE), affinity CE (ACE), frontal analysis (FA), Hummel-Dreyer method (HD), and vacancy peak (VP), have been used to determine affinity interactions [11][12][13]. Busch et al [13] have described the methodological differences between them.…”

Section: Five Ce Modesmentioning

confidence: 99%

“…Five CE modes including zone migration CE (CZE), affinity CE (ACE), frontal analysis (FA), Hummel-Dreyer method (HD), and vacancy peak (VP), have been used to determine affinity interactions [11][12][13]. Busch et al [13] have described the methodological differences between them. Table 1 summarizes the experimental details of these five modes.…”

Section: Five Ce Modesmentioning

confidence: 99%

“…In FA, there are two requirements: (i) the receptor and the complex have approximately the same mobility, and (ii) the mobility of the ligand differs sufficiently from the mobility of the complex. Also, in VP the receptor has an approximate mobility with the complex and HD requires that the ligand has a different mobility with respect to the receptor and the complex [13]. Among these five CE modes, HD and VP are analyte-consuming compared with the other modes but they are well suited for calculating the binding stoichiometry.…”

Section: Five Ce Modesmentioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

Section: Five Ce Modesmentioning

confidence: 99%

Section: Five Ce Modesmentioning

confidence: 99%

Section: Five Ce Modesmentioning

confidence: 99%

See 2 more Smart Citations

Recent advances in the study of biomolecular interactions by capillary electrophoresis

Ding

et al. 2004

Electrophoresis

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“…They are mostly based on the bound and free ligand separation in the condition of equilibrium. Among them there are methods like filtration, precipitation, chromatography which may disturb the mutual equilibrium of the constituents (Busch et al, 1997a). Capillary electrophoresis (CE) has become an important technique for studies of the nature and strength of biological interactions (Š olté s, 2004).…”

Section: Introductionmentioning

confidence: 99%

Binding of an Oxindole Alkaloid from Uncaria tomentosa to Amyloid Protein (Aβ1-40)

Frąckowiak

Bączek

Kaliszan

et al. 2006

Zeitschrift Für Naturforschung C

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The primary aim of this work was to determine the interactions of an oxindole alkaloid (mitraphylline) isolated from Uncaria tomentosa with -amyloid 1-40 (A 1-40 protein) applying the capillary electrophoresis (CE) method. Specifically the Hummel-Dreyer method and Scatchard analysis were performed to study the binding of oxindole alkaloids with A 1-40 protein. Prior to these studies extraction of the alkaloid of interest was carried out. Identification of the isolated alkaloid was performed by the use of thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) combined with electrospray ionization mass spectrometry (ESI-MS). The proposed approach was proved to be an efficient and accurate method for specific compound isolation and identification purposes. Moreover, analytical information from the CE approach can be considered as the valuable tool for binding constant determination. The binding constant of mitraphylline with A 1-40 protein determined by the Hummel-Dreyer method and Scatchard analysis equals K = 9.95 ¥ 10 5 m Ð1. The results obtained showed the significant binding of the tested compound with A 1-40 protein.These results are discussed and interpreted in the view of developing a strategy for identification of novel compounds of great importance in Alzheimer disease therapy.

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Study of interaction between drug enantiomers and serum albumin by capillary electrophoresis

1999

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The interaction between drugs and human serum albumin (HSA) was investigated by capillary electrophoresis (CE). It involves stereoselectivity, drug displacement and synergism effects. Under protein‐drug binding equilibrium, the unbound concentrations of drug enantiomers were measured by frontal analysis (FA). The stereoselectivity of verapamil (VER) binding to HSA was proved by the different free fractions of two enantiomers. In physiological pH (7.4, ionic strength 0.17 phosphate buffer) when 300 μM (±) VER were equilibrated with 500 μM HSA, the concentration of unbound S‐VER was about 1.7 times its antipode. The binding constants of two enantiomers, KR‐VER and KS‐VER, were 2670 and 850 M—1, respectively. However, no obvious stereoselective binding of propranolol (PRO) to HSA was observed. Trimethyl‐β‐cyclodextrin (45 mM) was used as a chiral selector in pH 2.5 phosphate buffer. Several drug systems were studied by the method. When ibuprofen (IBU) was added into VER‐HSA solution. R‐VER was partially displaced while S‐VER was not displaced at all. A binding synergism effect between bupivacaine (BUP) and verapamil was observed and further study suggested that verapamil and bupivacaine occupy different binding site of HSA (site II and site III, respectively).

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Comparison of five methods for the study of drug–protein binding in affinity capillary electrophoresis

Cited by 227 publications

References 26 publications

Recent advances in the study of biomolecular interactions by capillary electrophoresis

Recent advances in the study of biomolecular interactions by capillary electrophoresis

Binding of an Oxindole Alkaloid from Uncaria tomentosa to Amyloid Protein (Aβ1-40)

Study of interaction between drug enantiomers and serum albumin by capillary electrophoresis

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