Design and synthesis of affinity ligands and relation of their structure with adsorption of proteins

Ye, Long; Xu, Aizhang; Cheng, C. C.; Zhang, Li; Huo, Chenxi; Huang, Feiyun; Xu, Hong; Li, Rongxiu

doi:10.1002/jssc.201100606

Cited by 13 publications

(20 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The affinity media were prepared according to previously developed methods . Briefly, Sepharose 6B beads (100 g) were thoroughly washed with pure water at a 10% m/v ratio, drained and suspended in 50 mL of activating solution (1 M NaOH, 25% DMSO, and 10 mL epichlorohydrin), followed by incubation at 40°C for 2.5 h with shaking (Fig.…”

Section: Methodsmentioning

confidence: 99%

“…The beads were then washed with 50% v/v acetone to remove the unbound cyanuric chloride. The ninhydrin test was used to evaluate the binding of the amino groups : the purple color indicated free amino groups, whereas the color disappearance indicated the occurrence of the binding between cyanuric chloride and the amino group. A twofold excess of pABA dissolved in 2 M sodium carbonate was added to the dichlorotriazinylated Sepharose 6B beads.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Rapid and efficient one‐step purification of a serralysin family protease by using a p‐aminobenzamidine‐modified affinity medium

Wang

Lin

et al. 2017

J of Separation Science

View full text Add to dashboard Cite

The metalloproteinase MP belongs to the serralysin family, which is involved in important functions such as nutrient acquisition and infection pathogenesis. Serralysin proteases in highly purified form are commonly used at the industrial level with several purposes. In this study, we set up an efficient and rapid purification protocol for MP using a p-aminobenzamidine-modified affinity chromatography. The affinity medium was synthesized by using p-aminobenzamidine as affinity ligand immobilized via cyanuric chloride spacer to Sepharose 6B sorbent carrier. According to the adsorption analysis, the dissociation constant K and theoretical maximum adsorption Q of this medium were 24.2 μg/mL and 24.1 mg/g wet sorbent, respectively. The purity of MP was assessed by a high-performance liquid chromatography on a TSK3000SW column and sodium dodecyl sulfate polyacrylamide gel electrophoresis, revealing values of 98.7 and ∼98%, respectively. The specific activity of purified MP was 95.6 U/mg, which is similar to values obtained through traditional purification protocols. In conclusion, our protocol could be easily employed for the rapid isolation of MP with high purity, and could be implemented for other serralysin family proteases.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Rapid and efficient one‐step purification of a serralysin family protease by using a p‐aminobenzamidine‐modified affinity medium

Wang

Lin

et al. 2017

J of Separation Science

View full text Add to dashboard Cite

show abstract

“…Namely, the reactivity of cyanuric chloride (2,4,6-trichloro-striazine) towards amines together with its structural rigidity, the ease in generating molecular diversity, and the formation of nonfissile bonds with the substituents have prompted its extensive use for the generation of synthetic mimic bifunctional protein ligands [3,4,10]. Several biomimetic triazine-based ligands have been thereof successfully designed as stable synthetic analogues that replace natural biological ligands.…”

Section: Affinity Ligandsmentioning

confidence: 99%

Biomimetic Affinity Ligands for Protein Purification

Sousa

Taipa

2014

Methods in Molecular Biology

View full text Add to dashboard Cite

The development of sophisticated molecular modeling software and new bioinformatic tools, as well as the emergence of data banks containing detailed information about a huge number of proteins, enabled the de novo intelligent design of synthetic affinity ligands. Such synthetic compounds can be tailored to mimic natural biological recognition motifs or to interact with key surface-exposed residues on target proteins and are designated as "biomimetic ligands." A well-established methodology for generating biomimetic or synthetic affinity ligands integrates rational design with combinatorial solid-phase synthesis and screening, using the triazine scaffold and analogues of amino acids side chains to create molecular diversity.Triazine-based synthetic ligands are nontoxic, low-cost, highly stable compounds that can replace advantageously natural biological ligands in the purification of proteins by affinity-based methodologies.

show abstract

“…The structure-based design of the affinity ligand that serves as a specific inhibitor or substrate analogue is an efficient and commonly used approach in the affinity purification of enzymes [ 26 , 27 , 28 ]. An alkaline metalloprotease, MP (accession no.…”

Section: Introductionmentioning

confidence: 99%

Structure-Based Design and Synthesis of a New Phenylboronic-Modified Affinity Medium for Metalloprotease Purification

Wang²,

Xu³

et al. 2016

Marine Drugs

View full text Add to dashboard Cite

Metalloproteases are emerging as useful agents in the treatment of many diseases including arthritis, cancer, cardiovascular diseases, and fibrosis. Studies that could shed light on the metalloprotease pharmaceutical applications require the pure enzyme. Here, we reported the structure-based design and synthesis of the affinity medium for the efficient purification of metalloprotease using the 4-aminophenylboronic acid (4-APBA) as affinity ligand, which was coupled with Sepharose 6B via cyanuric chloride as spacer. The molecular docking analysis showed that the boron atom was interacting with the hydroxyl group of Ser176 residue, whereas the hydroxyl group of the boronic moiety is oriented toward Leu175 and His177 residues. In addition to the covalent bond between the boron atom and hydroxyl group of Ser176, the spacer between boronic acid derivatives and medium beads contributes to the formation of an enzyme-medium complex. With this synthesized medium, we developed and optimized a one-step purification procedure and applied it for the affinity purification of metalloproteases from three commercial enzyme products. The native metalloproteases were purified to high homogeneity with more than 95% purity. The novel purification method developed in this work provides new opportunities for scientific, industrial and pharmaceutical projects.

show abstract

Design and synthesis of affinity ligands and relation of their structure with adsorption of proteins

Cited by 13 publications

References 23 publications

Rapid and efficient one‐step purification of a serralysin family protease by using a p‐aminobenzamidine‐modified affinity medium

Rapid and efficient one‐step purification of a serralysin family protease by using a p‐aminobenzamidine‐modified affinity medium

Biomimetic Affinity Ligands for Protein Purification

Structure-Based Design and Synthesis of a New Phenylboronic-Modified Affinity Medium for Metalloprotease Purification

Contact Info

Product

Resources

About