Affinity capillary electrophoresis

Rippel, Géza; Corstjens, Hugo; Billiet, H.A.H.; Frank, Johannes

doi:10.1002/elps.1150181208

Cited by 59 publications

(35 citation statements)

References 82 publications

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“…Among the existing CE methods developed for measuring binding constants (K b ) [11], affinity capillary electrophoresis (ACE) has been shown to be quite simple and effective [6,10,12,13]. Other advantages include versatility, high speed, small sample consumption, and low operational costs.…”

Section: Introductionmentioning

confidence: 99%

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Comparative studies of the interaction between ferulic acid and bovine serum albumin by ACE and surface plasmon resonance

Zhang

et al. 2007

Electrophoresis

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Affinity capillary electrophoresis (ACE) was used to study the interaction between ferulic acid (FA) and BSA. The interaction between FA and BSA was facilitated by injecting FA into a BSA-containing running buffer. Both mobility ratio and mobility shift assays were performed to deduce the binding constant (K(b)). However, the K(b )value obtained with the mobility ratio assay was only approximately 20% of that extracted from the mobility shift assay. The former assay yielded a K(b) value (5.6 +/- 0.4 x 10(4) M(-1)), which compares well with the result obtained with surface plasmon resonance (SPR) (5.1 +/- 0.6 x 10(4) M(-1)). The discrepancy between the mobility ratio and mobility shift assays suggests that the data extrapolation from the mobility ratio should be more reliable for cases when both changes in the EOF and viscosity of the running buffer are important. The work demonstrates that ACE, a solution-based technique, and SPR, a technique addressing interfacial processes, are highly complementary to each other and the comparative studies are confirmatory and allow binding constants to be accurately determined.

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Section: Introductionmentioning

confidence: 99%

“…The use of CE [1][2][3][4] to study biological interactions [5][6][7][8][9][10] is becoming increasingly popular. Among the existing CE methods developed for measuring binding constants (K b ) [11], affinity capillary electrophoresis (ACE) has been shown to be quite simple and effective [6,10,12,13].…”

Section: Introductionmentioning

confidence: 99%

Comparative studies of the interaction between ferulic acid and bovine serum albumin by ACE and surface plasmon resonance

Zhang

et al. 2007

Electrophoresis

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“…Since CE features high efficiency, quick analysis, and microvolume (usually nanoliter), it rapidly became popular in analytical fields. In general, CE binding studies involve either measuring the solute mobility change caused by the addition of another entity to the system, which is called affinity capillary electrophoresis [1,2], or estimating the unbound concentration of the analyte in a mixed solution. Affinity capillary electrophoresis (ACE) is widely used in binding studies [3,4], but it usually has some drawbacks [5,6], such as strong protein UV absorption, and poor reproducibility of migration time.…”

Section: Introductionmentioning

confidence: 99%

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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“…IACE (II, III) can be considered an equilibrium technique [43], but it does not suffer from disturbances (adsorption, solubility, recovery) related to two-phase systems such as LC.…”

Section: Theoretical Aspects Of Iacementioning

confidence: 99%