Affinity Chromatography as a Tool for Quantification of Interactions Between Drug Molecules and Their Protein Targets

Drączkowski, Piotr; Matosiuk, Dariusz; Jóźwiak, Krzysztof

doi:10.5772/36282

Cited by 3 publications

(2 citation statements)

References 84 publications

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“…The nucleotide affinity to their targets can be influenced by selection conditions. In some cases, binding and washing conditions, such as target molecule concentration, buffer composition, time, and volume, are modified in later SELEX cycles to obtain aptamers with high affinity and specificity [29,39].…”

Section: Separation Of Amplified Aptamersmentioning

confidence: 99%

New Insights into Aptamers: An Alternative to Antibodies in the Detection of Molecular Biomarkers

Domsicova,

Korcekova,

Poturnayova

et al. 2024

IJMS

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Aptamers are short oligonucleotides with single-stranded regions or peptides that recently started to transform the field of diagnostics. Their unique ability to bind to specific target molecules with high affinity and specificity is at least comparable to many traditional biorecognition elements. Aptamers are synthetically produced, with a compact size that facilitates deeper tissue penetration and improved cellular targeting. Furthermore, they can be easily modified with various labels or functional groups, tailoring them for diverse applications. Even more uniquely, aptamers can be regenerated after use, making aptasensors a cost-effective and sustainable alternative compared to disposable biosensors. This review delves into the inherent properties of aptamers that make them advantageous in established diagnostic methods. Furthermore, we will examine some of the limitations of aptamers, such as the need to engage in bioinformatics procedures in order to understand the relationship between the structure of the aptamer and its binding abilities. The objective is to develop a targeted design for specific targets. We analyse the process of aptamer selection and design by exploring the current landscape of aptamer utilisation across various industries. Here, we illuminate the potential advantages and applications of aptamers in a range of diagnostic techniques, with a specific focus on quartz crystal microbalance (QCM) aptasensors and their integration into the well-established ELISA method. This review serves as a comprehensive resource, summarising the latest knowledge and applications of aptamers, particularly highlighting their potential to revolutionise diagnostic approaches.

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Section: Separation Of Amplified Aptamersmentioning

confidence: 99%

New Insights into Aptamers: An Alternative to Antibodies in the Detection of Molecular Biomarkers

Domsicova,

Korcekova,

Poturnayova

et al. 2024

IJMS

View full text Add to dashboard Cite

show abstract

“…Many forces are involved in the intermolecular association, including hydrophobic, van der Waals, or stacking interactions between aromatic amino acids, hydrogen bonding, and electrostatic forces. 26 Besides, there are many degrees of freedom (DOF) as well as insufficient knowledge of the effect of solvent on the binding association. The process of docking a ligand to a binding site tries to mimic the natural course of interaction of the ligand and its receptor via the lowest energy pathway.…”

Section: Optimization Of the Adsorbent And Chromatographic Parametersmentioning

confidence: 99%

Molecular docking of metal ion immobilized ligands to proteins in affinity chromatography

Salha

Andaç

Deni̇zli̇

2020

J of Molecular Recognition

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Immobilized metal ion affinity chromatography (IMAC) has become a widespread analytical and preparative separation method for therapeutic proteins, peptides nucleic acids, hormones, and enzymes. N‐Methacryloyl‐l‐histidine Methyl Ester (MAH) monomer is recently used as a synthesized affinity ligand in IMAC. It is capable of chelating with many transition metal ions such as Zn2+, Ni2+, and Cu2+ ions through its histidine residue. In this way, proteins can bind selectively to these immobilized metal ions through MAH as a ligand in affinity chromatography. In this study, we applied the computational docking method on the interactions that occur between the MAH monomer and its complexes with Zn2+ ions as ligands and protein molecules as targets. MAH monomer was drawn and created using the Avogadro software as an optimization tool. Human insulin (Ins) molecule and horse heart cytochrome C (Cyt C) were selected as target proteins to interact with MAH monomer as affinity ligand. Automated docking software AutoDock v4.2 was used for docking of MAH monomer to Ins and Cyt C, respectively. The affinity ligand complexes with Zn2+ ions bound to one, two, and three moles of MAH were studied and compared separately. The lowest binding energies of Ins and Cyt C proteins in 1:1 mol ratio of MAH‐Zn2+ were found as (−4.14) and (−4.92) kcal/mol, respectively.

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Penetrable silica microspheres for immobilization of bovine serum albumin and their application to the study of the interaction between imatinib mesylate and protein by frontal affinity chromatography

Zhao

et al. 2015

Anal Bioanal Chem

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In the current study, novel featured silica, named penetrable silica, simultaneously containing macropores and mesopores, was immobilized with bovine serum albumin (BSA) via Schiff base method. The obtained BSA-SiO2 was employed as the high-performance liquid chromatographic (HPLC) stationary phase. Firstly, D- and L-tryptophan were used as probes to investigate the chiral separation ability of the BSA-SiO2 stationary phase. An excellent enantioseparation factor was obtained up to 4.3 with acceptable stability within at least 1 month. Next, the BSA-SiO2 stationary phase was applied to study the interaction between imatinib mesylate (IM) and BSA by frontal affinity chromatography. A single type of binding site was found for IM with the immobilized BSA, and the hydrogen-bonding and van der Waals interactions were expected to be contributing interactions based on the thermodynamic studies, and this was a spontaneous process. Compared to the traditional silica for HPLC stationary phase, the proposed penetrable silica microsphere possessed a larger capacity to bond more BSA, minimizing column overloading effects and enhancing enantioseparation ability. In addition, the lower running column back pressure and fast mass transfer were meaningful for the column stability and lifetime. It was a good substrate to immobilize biomolecules for fast chiral resolution and screening drug-protein interactions.

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Affinity Chromatography as a Tool for Quantification of Interactions Between Drug Molecules and Their Protein Targets

Cited by 3 publications

References 84 publications

New Insights into Aptamers: An Alternative to Antibodies in the Detection of Molecular Biomarkers

New Insights into Aptamers: An Alternative to Antibodies in the Detection of Molecular Biomarkers

Molecular docking of metal ion immobilized ligands to proteins in affinity chromatography

Penetrable silica microspheres for immobilization of bovine serum albumin and their application to the study of the interaction between imatinib mesylate and protein by frontal affinity chromatography

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