Determination of dissociation constants of complexes of trypsin and its low molecular weight inhibitors by affinity chromatography in zonal and frontal analysis arrangement

Malaníková, M.; Turková, J.

doi:10.1007/bf02996745

Cited by 13 publications

(1 citation statement)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3c , and we use the most likely value in the histogram to be the value of K D , which is found to be 34.7 μM. Three repeated runs were performed, and the averaged value agrees with the reported K D from literatures 27 28 ( Table 1 ). The minor difference could be due to different buffers and pH value in different experiments (we have used 5 mM Tris-HCl buffer at pH = 7.4).…”

supporting

confidence: 71%

Transient Induced Molecular Electronic Spectroscopy (TIMES) for study of protein-ligand interactions

Zhang

Han

et al. 2016

Sci Rep

View full text Add to dashboard Cite

We present a method, Transient Induced Molecular Electronic Spectroscopy (TIMES), to detect protein-ligand interactions without any protein engineering or chemical modification. We developed a physics model for the TIMES signal and mathematically formulated the problem to attain physical insight of protein-ligand interactions without any disturbances by molecular probes, fluorescent labels, or immobilization of molecules. To demonstrate the functionality of this method, we have used the TIMES signals to find the dissociation constants for the affinity of reactions, the shear-stress dependent adsorption time of molecules on surface, and other interesting features of protein-ligand interaction in native conditions. As a unique tool, TIMES offers a simple and effective method to investigate fundamental protein chemistry and drug discoveries.

show abstract

supporting

confidence: 71%

Transient Induced Molecular Electronic Spectroscopy (TIMES) for study of protein-ligand interactions

Zhang

Han

et al. 2016

Sci Rep

View full text Add to dashboard Cite

show abstract

An affinity adsorbent derived from aminopropyl silica for serine protease chromatography

Chang

Lee

1994

J of Chemical Tech & Biotech

View full text Add to dashboard Cite

A silica‐based adsorbent for affinity chromatography of serine protease was prepared by bonding p‐aminobenzamidine (pABZ) to aminopropyl silica. Silanization of silica with both γ‐aminopropyltrimethoxysilane and γ‐amino‐propyltriethoxysilane under anhydrous conditions led to a monolayer density of primary amino groups. The aminopropyl silica was converted to primary hydroxylcontaining silica via diazotization, and consequently activated with p‐nitrophenyl chloroformate and immobilized with ligand pABZ. The resultant silica–pABZ has a ligand density of 13.8 μmol g−1. Pyridine was indicated by the data to increase the reactivity of the chloroformate toward the hydroxyl group more efficiently than 4‐dimethylaminopyridine. Two stages of adsorption were found in batch adsorption of trypsin with an equilibrium adsorption isotherm of the Langmuir type. When the chromatographic column packed with this silica–p ABZ was operated under a higher flow rate (2·33 cm3 min−1) and with 3 g dm−3 of crude urokinase as the influence, the yield was 55%. Both the flow rate and the concentration of the crude protein were shown, by measuring the dynamic binding capacity from chromatographic experiments, to be the factors which influenced the chromatographic efficiency.

show abstract

Theoretical Aspects of Quantitative Affinity Chromatography

Vidal-Madjar¹,

Jaulmes²

1992

Theoretical Advancement in Chromatography and Related Separation Techniques

View full text Add to dashboard Cite

Determination of dissociation constants of complexes of trypsin and its low molecular weight inhibitors by affinity chromatography in zonal and frontal analysis arrangement

Cited by 13 publications

References 15 publications

Transient Induced Molecular Electronic Spectroscopy (TIMES) for study of protein-ligand interactions

Transient Induced Molecular Electronic Spectroscopy (TIMES) for study of protein-ligand interactions

An affinity adsorbent derived from aminopropyl silica for serine protease chromatography

Theoretical Aspects of Quantitative Affinity Chromatography

Contact Info

Product

Resources

About