Immobilized Nicotinic Receptor Stationary Phase for On-Line Liquid Chromatographic Determination of Drug–Receptor Affinities

Zhang, Yanxiao; Xiao, Yingxian; Kellar, Kenneth J.; Wainer, Irving W.

doi:10.1006/abio.1998.2828

Cited by 86 publications

(64 citation statements)

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“…This study stimulated Wainer's group to approach the immobilization process of nicotinic acetylcholine receptors (nAChRs) on lipid monolayers [13]. Interestingly α 3 β 4 nAChRs were also successfully immobilized on the immobilized artificial membrane (IAM) stationary phase, developed by Pidgeon et al [14] and currently available from Regis Technologies, Figure 2.…”

Section: Cellular Membrane Affinity Chromatography Columns -A Short Hmentioning

confidence: 93%

“…Interestingly α 3 β 4 nAChRs were also successfully immobilized on the immobilized artificial membrane (IAM) stationary phase, developed by Pidgeon et al [14] and currently available from Regis Technologies, Figure 2. Studies by the Wainer group showed that CMAC prepared with the use of IAM particles were characterized by better reproducibility compared to liposomal columns [13]. Since the successful immobilization of nAChRs on the IAM stationary phase, multiple transmembrane proteins have been used to prepare CMACs with IAM.PC (immobilized artificial membrane phosphatidylcholine).…”

Section: Cellular Membrane Affinity Chromatography Columns -A Short Hmentioning

confidence: 99%

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Cellular membrane affinity chromatography (CMAC) in drug discovery from complex natural matrices

Stephen¹,

Omri²,

Cieśla

2018

ADMET DMPK

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Secondary plant metabolites are evolutionary-designed molecules that interact with multiple biological targets in human organisms. Identification of pharmacologically active phytochemicals is usually a time consuming and costly process. Cellular membrane affinity chromatography (CMAC) allows the detection of secondary metabolites present in complex natural matrices, e.g. plant extracts and their interactions with the immobilized fully-functional transmembrane proteins. After the isolation process of the binding compounds, CMAC columns can be used to study the binding process between the potential new ligands and the immobilized transmembrane protein target. The following parameters can be determined using CMAC columns: binding affinity (Kd), association rate constant (kon), dissociation rate constant (koff) and the equilibrium constant for complex formation (K). This review summarizes the preparation steps and the use of CMAC columns in the drug discovery process of new potential drug leads present in complex natural matrices.

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Section: Cellular Membrane Affinity Chromatography Columns -A Short Hmentioning

confidence: 93%

Section: Cellular Membrane Affinity Chromatography Columns -A Short Hmentioning

confidence: 99%

Cellular membrane affinity chromatography (CMAC) in drug discovery from complex natural matrices

Stephen¹,

Omri²,

Cieśla

2018

ADMET DMPK

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“…23,[64][65][66][67][68][69][70] No final da década de 90, IMERs contendo receptores e transportadores trans-membrana imobilizados foram desenvolvidos para triagem on-line de misturas complexas no desenvolvimento racional de fármacos. Os receptores de membrana são mediadores das etapas iniciais da atividade dos fármacos.…”

Section: Década De 90unclassified

“…Muitas vezes o aumento da estabilidade, da seletividade e da atividade da enzima é obtido através da combinação de diferentes técnicas de imobilização e da seleção apropriada do suporte. 16,17,120 A Tabela 5 lista uma variedade de suportes cromatográficos e diversas aplicações. Assim, as vantagens e desvantagens entre os métodos e suportes devem ser consideradas em cada caso específico.…”

Section: Suportes Cromatográficosunclassified

“…A imobilização de enzimas a suportes cromatográficos apresenta diversas vantagens, sobre a utilização de enzimas livres em solução, entre as quais destacamos: a utilização de pequenos volumes de amostra (pL-nL); aumento do tempo de vida e da estabilidade da enzima em relação à temperatura, aos solventes orgânicos e à variação de pH sem perda considerável da atividade catalítica; reutilização; pequeno manuseio da amostra, evitando contaminações; fácil separação da enzima dos produtos da reação. 16,17 Essas vantagens são úteis nos estudos enzimáticos on-line, onde os produtos da reação enzimática e os inibidores são detectados diretamente por UV-Vis, fluorescência ou espectrometria de massas. 18 Existem duas principais classes de aplicações analíticas dos biorreatores enzimáticos ou IMER.…”

Section: Introductionunclassified

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Imobilização de enzimas em suportes cromatográficos: uma ferramenta na busca por substâncias bioativas

Cardoso¹,

Moraes²,

Cass³

2009

Quím. Nova

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Recebido em 5/10/07; aceito em 11/7/08; publicado na web em 2/12/08 IMMOBILIZATION OF THE ENZYMES ON CHROMATOGRAPHIC SUPPORTS: A TOOL TO RESEARCH OF INHIBITOR COMPOUNDS. The development and characterization of bioreactors or IMER (immobilized enzyme reactors) as research tools are important in the scope of medicinal chemistry and constitute an alternative for the rational development of drugs. This approach does not require highly purified enzymes or a great amount of protein, but increase the enzymatic stability against heat, organic solvents and pH, without too much loss of catalyst activity. Immobilized enzyme reactors (IMER) can be used for the accomplishment of high efficiency screening on-line and, thus inhibitors can be quickly identified. Here, we emphasize the development of IMER by use of different methods of immobilization and chromatographic supports. Their applications, in different areas of research, are also fully discussed.

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Protein–Drug Interactions

Oravcová¹,

Lindner

2000

Encyclopedia of Analytical Chemistry

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Noncovalent (reversible) drug–protein binding interactions are an integral part of intermolecular recognition processes taking place in a cell or organ environment. Characterization of basic constraints of target protein binding site(s) (affinity, stoichiometry, specificity, saturability and stereoselectivity) for particular ligand(s) is an essential output of many different drug–protein binding studies. Nonlinear and stereoselective binding phenomena, including drug–drug (or enantiomer–enantiomer) interactions and their e.g. genetically or pathophysiologically determined variability, can be predicted in this way. In general, the methods used for drug–protein binding studies can be divided into separation (nonspectroscopic) and spectroscopic methods. Separation methods are particularly suitable for the quantitative determination of drug binding, whereas spectroscopic techniques are able to characterize the qualitative features of drug–protein complexes (e.g. conformational status/changes). The primary objective was to summarize the most relevant trends in separation techniques [affinity high‐performance liquid chromatography (HPLC) and affinity capillary electrophoresis (ACE)] employed in the framework of drug–protein/biopolymer binding assays and to compare them with so‐called conventional approaches (equilibrium dialysis (ED), ultrafiltration). Focused attention has been paid particularly to HPLC and capillary electrophoresis (CE) methods designed for drug–protein studies when so‐called “dynamic” and “secondary” equilibria are involved and created. A common feature of this strategy is to use binding protein(s) or interacting drug(s) without their covalent immobilization as mobile phase or background electrolyte (BGE) (buffer) additives. This offered a new, elegant tool for the separation of “bound” and “free” forms of interacting species in a “miniaturized free solution system” operating under nearly physiological conditions. Undoubtedly, it permits simplification of the experimental protocols where it is crucial to perform repetitive analyses under “the same starting conditions”, as e.g. the occupancy of binding sites is concerned.

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Immobilized Nicotinic Receptor Stationary Phase for On-Line Liquid Chromatographic Determination of Drug–Receptor Affinities

Cited by 86 publications

References 10 publications

Cellular membrane affinity chromatography (CMAC) in drug discovery from complex natural matrices

Cellular membrane affinity chromatography (CMAC) in drug discovery from complex natural matrices

Imobilização de enzimas em suportes cromatográficos: uma ferramenta na busca por substâncias bioativas

Protein–Drug Interactions

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