Use of a partial-filling technique in affinity capillary electrophoresis for determining binding constants of ligands to receptors

Heintz, Joseph; Hernandez, Monica C.; Gomez, Frank A.

doi:10.1016/s0021-9673(99)00278-2

Cited by 71 publications

(64 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…But it seems not easy to find two markers which do not interact with the receptor and ligand. A two-step procedure with on-column ligand synthesis and partically filling ACE has been described [128,129]. This approach can couple on-line reaction with the separation.…”

Section: Small Molecule-small Molecule Interactionmentioning

confidence: 99%

Recent advances in the study of biomolecular interactions by capillary electrophoresis

Ding

et al. 2004

Electrophoresis

View full text Add to dashboard Cite

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.

show abstract

Section: Small Molecule-small Molecule Interactionmentioning

confidence: 99%

Recent advances in the study of biomolecular interactions by capillary electrophoresis

Ding

et al. 2004

Electrophoresis

View full text Add to dashboard Cite

show abstract

“…So far, only a few reports on determination of affinity constants using the partial-filling technique are available [27][28][29][30]. The main reasons for this weak interest (so far) might be technical shortcomings of the instrument, difficulties in the preparation or availability of suitable capillaries, and complicated data treatment.…”

Section: General Aspectsmentioning

confidence: 99%

Determination of protein‐ligand affinity constants from direct migration time in capillary electrophoresis

Nilsson¹,

Harang

Bergström³

et al. 2004

Electrophoresis

View full text Add to dashboard Cite

A simple method to calculate dissociation constants for protein-ligand interactions by partial-filling capillary electrophoresis is demonstrated. The method uses raw migration time data for the ligand and needs only additional information about capillary inner radius and the absolute amount of protein loaded. A theoretical study supported by experimental data also demonstrates that the retention of analyte in affinity capillary electrophoresis (ACE) using the partial-filling technique depends linearly on the absolute amount of selector added but is independent of both selector zone length and selector mobility. Factors such as field strength and electroosmotic flow are also cancelled out if they are kept constant. The theory is confirmed and the usefulness of the method is demonstrated by enantioseparations using alpha-acid glycoprotein (AGP) and cellulase (Cel 7A) as chiral selectors.

show abstract

“…The use of mobility ratios (M) was found to give more precise estimates of binding constants when large changes in the EOF are observed. [41][42][43] Mobility ratios are calculated as:…”

Section: Determination Of the Association Constants Between Proteins mentioning

confidence: 99%

“…This method has been widely applied to separation, enantioseparation and determination of association constants. [28][29][30][31][32] Human serum albumin has been used as a chiral selector to separate many drugs in CE. 19,26,33,34 However, the chiral separation of mexiletine and chlorpheniramine with HSA as selector has not been reported.…”

Section: Introductionmentioning

confidence: 99%

Enantiomeric Separation of Basic Drugs with Partially Filled Serum Albumin as Chiral Selector in Capillary Electrophoresis

Chen

2004

ANAL. SCI.

View full text Add to dashboard Cite

A reliable method is presented for the chiral separation of three basic drugs (mexiletine, chlorpheniramine and propranolol) with serum albumins (human and porcine, HSA and PSA) as chiral selectors by capillary electrophoresis in combination with the partial filling technique. Based on the systematic optimization of operation variables, the chiral separation of mexiletine, chlorpheniramine and propranolol was achieved in the pH 7.4 phosphate buffer by using HSA, PSA and PSA as selectors, respectively. The chiral recognition ability of HSA and PSA was compared. HSA and PSA show a different chiral recognition ability for each of the three drugs. In addition, the association constants between enantiomeric drugs and proteins were determined to be 2.00 and 3.80 × 10 2 M -1 for mexiletine and HSA, 0.59 and 1.12 × 10 3 M -1 for chlorpheniramine and PSA, and 0.87 and 1.42 × 10 3 M -1 for propranolol and PSA. The method for the chiral separation and determination of association constants possesses the advantages of simple performance, effective avoiding of the interference of the UV detection from protein, and lowering of the reagent consumption.

show abstract

Use of a partial-filling technique in affinity capillary electrophoresis for determining binding constants of ligands to receptors

Cited by 71 publications

References 57 publications

Recent advances in the study of biomolecular interactions by capillary electrophoresis

Recent advances in the study of biomolecular interactions by capillary electrophoresis

Determination of protein‐ligand affinity constants from direct migration time in capillary electrophoresis

Enantiomeric Separation of Basic Drugs with Partially Filled Serum Albumin as Chiral Selector in Capillary Electrophoresis

Contact Info

Product

Resources

About