Estimation of Receptor–Ligand Interactions by the Use of a Two-Marker System in Affinity Capillary Electrophoresis

Mito, Erica; Zhang, Ying; Esquivel, Sally; Gomez, Frank A.

doi:10.1006/abio.2000.4536

Cited by 36 publications

(30 citation statements)

References 35 publications

(35 reference statements)

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“…In this form of analysis K b is estimated using a dual-marker form of analysis, which we term the relative migration time ratio (RMTR) (Eq. 1) [25].…”

Section: Resultsmentioning

confidence: 98%

On‐column derivatization of the antibiotics teicoplanin and ristocetin coupled to affinity capillary electrophoresis

2003

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Binding constants between the glycopeptides teicoplanin (Teic) and ristocetin (Rist) and their derivatives to D-Ala-D-Ala terminus peptides were determined by on-column receptor synthesis coupled to partial-filling affinity capillary electrophoresis (PFACE) or affinity capillary electrophoresis (ACE). In these techniques, the column is first partially filled with increasing concentrations of D-Ala-D-Ala terminus peptides. This is followed by plugs of buffer, antibiotic and two noninteracting standards, and acetic and/or succinic anhydride (and buffer in the case of ACE). The order of the reagent plugs containing the antibiotic and anhydride varies with the charge of the glycopeptide. Upon electrophoresis, the antibiotic reacts with the anhydride yielding a derivative of Teic or Rist. Continued electrophoresis results in the overlap of the derivatized antibiotic and the plug of D-Ala-D-Ala peptide. Analysis of the change in the relative migration time ratio (RMTR) of the new glycopeptide relative to the standards, as a function of the concentration of the D-Ala-D-Ala ligand yields a value for the binding constant K(b). The techniques described here can be used to assess how the derivatization of drugs alters their affinities for target molecules.

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“…In this form of analysis K b is estimated using a dual-marker form of analysis, which we term the relative migration time ratio (RMTR) (Eq. 1) [25].…”

Section: Resultsmentioning

confidence: 98%

On‐column derivatization of the antibiotics teicoplanin and ristocetin coupled to affinity capillary electrophoresis

2003

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Section: Use Of Chemometric Methodology In Optimizing Conditions For mentioning

confidence: 99%

“…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…”

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confidence: 99%

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

2008

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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

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“…ET-1, the ET-1 fragment ET-1 (16)(17)(18)(19)(20)(21) containing 16-21 residues [26,27], and the cyclic peptide antagonists BQ123 [28] and JKC302 [29,30] were synthesized by using an ABI 433A peptide synthesizer (ABI, Foster City, CA, USA). We also prepared the samples of non-peptide endothelin receptor antagonists SB209670 [31,32], JMF310 [33], and YHK891 [34].…”

Section: Samples and Reagentsmentioning

confidence: 99%

Capillary electrophoresis using immobilized whole cells with overexpressed endothelin receptor for specific ligand screening

Yu¹,

Tseng²,

Fang³

et al. 2004

Electrophoresis

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A new capillary electrophoresis method using immobilized cells as the stationary phase has been developed. The power of this method is demonstrated by the separation and identification of endothelin antagonists on a capillary column coated by the transfected Chinese hamster ovary (CHO) cells with overexpressing endothelin receptors. The screening results are validated by functional assays suppressing the increase of intracellular calcium concentration induced by endothelin-1. Instead of making efforts in isolating protein receptors, the easily prepared whole-cell capillary column provides a superior tool on the basis of ligand/receptor affinity for a rapid screening of potent drug candidates from compound libraries.

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Estimation of Receptor–Ligand Interactions by the Use of a Two-Marker System in Affinity Capillary Electrophoresis

Cited by 36 publications

References 35 publications

On‐column derivatization of the antibiotics teicoplanin and ristocetin coupled to affinity capillary electrophoresis

On‐column derivatization of the antibiotics teicoplanin and ristocetin coupled to affinity capillary electrophoresis

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

Capillary electrophoresis using immobilized whole cells with overexpressed endothelin receptor for specific ligand screening

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