Flow‐through partial‐filling affinity capillary electrophoresis can estimate binding constants of neutral ligands to receptors <b><i>via</i></b> a competitive assay technique

Kaddis, John; Mito, Erica; Heintz, Joseph; Plazas, A.; Gomez, Frank A.

doi:10.1002/elps.200390129

Cited by 33 publications

(36 citation statements)

References 32 publications

(18 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…, and its on-column modified derivative [62]. Other new modifications of PF-CAE involve flowthrough PF-CAE, competitive flow-through PF-CAE, and multiple-step ligand injection PF-CAE [129,130]. Optimization of conditions for flow-through PF-CAE, including the determination of minimal injection time of ligand required to estimate binding constants of dipeptide D-Ala-D-Ala ligand to glycopeptide antibiotic, vancomycin, has been recently reported [131].…”

Section: Affinity Electrophoresismentioning

confidence: 97%

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

2006

View full text Add to dashboard Cite

The article gives a comprehensive review on the recent developments in the applications of high-performance capillary electromigration methods, zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography, to analysis, preparation, and physicochemical characterization of peptides. The article presents new approaches to the theoretical description and experimental verification of electromigration behavior of peptides, covers the methodological aspects of capillary electroseparations of peptides, such as rational selection of separation conditions, sample preparation, suppression of peptide adsorption, new developments in individual separation modes, and new designs of detection systems. Several types of applications of capillary electromigration methods to peptide analysis are presented: conventional qualitative and quantitative analysis, purity control, determination in biomatrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid and sequence analysis, and peptide mapping of proteins. Some examples of micropreparative peptide separations are given and capabilities of capillary electromigration techniques to provide important physicochemical characteristics of peptides are demonstrated.

show abstract

Section: Affinity Electrophoresismentioning

confidence: 97%

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

2006

View full text Add to dashboard Cite

show abstract

“…In this study, Factors, levels, and a target K bo value of 2.50610 6 M 21 were selected based on our previous univariate work on FTPFACE and with the CAB-charged arylsulfonamide system [9]. This K bo value is in agreement with previous ACE studies and in studies using other assay techniques [8,33].…”

Section: Experimental Design and Optimization Proceduresmentioning

confidence: 76%

“…During electrophoresis the zones of samples overlap within the capillary and an equilibrium is established prior to the point of detection. An extension of PFACE is flow-through PFACE (FTPFACE) [9,11,19]. In this technique, the capillary column is partially filled with an even smaller plug of ligand than that used in PFACE (Fig.…”

Section: Introductionmentioning

confidence: 99%

Use of chemometric methodology in optimizing conditions for competitive binding partial filling affinity capillary electrophoresis

2008

Self Cite

View full text Add to dashboard Cite

Use of chemometric methodology in optimizing conditions for competitive binding partial filling affinity capillary electrophoresisThis work expands the knowledge of the use of chemometric response surface methodology (RSM) in optimizing conditions for competitive binding partial filling ACE (PFACE). Specifically, RSM in the form of a Box-Behnken design was implemented in flow-through PFACE (FTPFACE) ACE has been shown to be a versatile microanalytical technique to estimate affinity constants, and has emerged as a useful and sensitive method for studying bimolecular noncovalent interactions and for determining binding and dissociation constants of formed complexes. The first reports detailing the use of ACE to measure affinity parameters between biological species were published in the early 1990s [3][4][5][6][7]. Since these informative studies, a multitude of other interactions including protein-ligand, peptide-peptide, proteinpeptide, protein-antibody, polymer-peptide, and antibodyantigen have been examined successfully using ACE .ACE differentiates between bound and unbound receptor (R) as a function of free ligand (L) concentration only when the R-L complexation yields a sizable difference in mass or charge-to-mass ratio. In a typical ACE experiment, a sample of receptor and noninteracting markers are reacted with an increasing concentration of ligand in a running buffer, thereby, causing a shift in the migration of the receptor peak. Subsequent analysis of these changes in migration time yields a value for K b [20].To minimize the amount of sample needed in an ACE assay, partial filling techniques in ACE were developed. In PFACE, the capillary is partially filled with ligand (or receptor) Correspondence: Dr. Frank A. Gomez, Department of Chemistry and Biochemistry, California State University, Los Angeles, CA, USA E-mail: fgomez2@calstatela.edu Fax: 11-323-343-6490 Abbreviations: CAB, carbonic anhydrase B; FTPFACE, flowthrough partial filling ACE; HHM, horse heart myoglobin; MO, mesityl oxide; RMTR, relative migration time ratio; RSM, response surface methodology

show abstract

“…However, some other studies focus on increasing the separation efficiency [19][20][21][22][23]. In a recent study, the partial filling technique and competitive binding have been combined [14].…”

Section: Introductionmentioning

confidence: 99%

Determination of dissociation constants by competitive binding in partial filling capillary electrophoresis

Nilsson¹,

Johansson

Isaksson³

2004

Electrophoresis

View full text Add to dashboard Cite

The determination of dissociation constands (K(d)) by competitive ligand binding in partial filling capillary electrophoresis is demonstrated. Two different strategies were applied, one of which only uses a single reporter ligand and a more elaborated one which suppresses systemic disturbances by using a racemic mixture as reporter. The dissociation constants obtained by both alternatives were virtually identical and in good agreement with those previously reported.

show abstract

Flow‐through partial‐filling affinity capillary electrophoresis can estimate binding constants of neutral ligands to receptors via a competitive assay technique

Cited by 33 publications

References 32 publications

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

Recent developments in capillary electrophoresis and capillary electrochromatography of peptides

Use of chemometric methodology in optimizing conditions for competitive binding partial filling affinity capillary electrophoresis

Determination of dissociation constants by competitive binding in partial filling capillary electrophoresis

Contact Info

Product

Resources

About