Mild and cost-effective green fluorescent protein purification employing small synthetic ligands

Pina, Ana Sofia; Dias, Ana; Üstok, Fatma Işık; Khoury, Graziella El; Fernandes, Cláudia S.M.; Branco, Ricardo; Lowe, Christopher R.; Roque, Ana C. A.

doi:10.1016/j.chroma.2015.09.036

Cited by 12 publications

(3 citation statements)

References 32 publications

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“…The ligand A4C7 is another example of an Ugi ligand developed to bind Green Fluorescent Protein (GFP). The affinity ligand showed good results in purifying GFP from crude extracts obtaining 94% yield and purity under very mild elution conditions (0.1M glycine pH 9) [79]. Furthermore, the ability to purify GFP-tagged proteins was demonstrated with a WW domain tagged to GFP with purities and yields of 92% and 68%, respectively, hence showing the dual applicability of A4C7.…”

Section: Biomimetic Ligandsmentioning

confidence: 96%

“…It involves a reaction between an amine, an aldehyde or ketone, an isonitrile and a carboxylic acid which results in a peptidomimetic product. The greater diversity allowed in this multi-component reaction enabled the successful application of this scaffold for novel adsorbents towards glycoproteins, erythropoietin (EPO), and affinity tags [75][76][77][78][79][80][81]. The Ugi scaffold has been applied to the generation of protein G biomimetic.…”

Section: Biomimetic Ligandsmentioning

confidence: 99%

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Rational design of affinity ligands for bioseparation

Matos

Pina

Roque

2020

Journal of Chromatography A

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Section: Biomimetic Ligandsmentioning

confidence: 96%

Section: Biomimetic Ligandsmentioning

confidence: 99%

Rational design of affinity ligands for bioseparation

Matos

Pina

Roque

2020

Journal of Chromatography A

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“…C-C and C-N bond-forming U-4CR can constitute a scaffold providing enormous molecular diversity. U-4CR protocol has been utilized in various fields of chemistry including separation science where it has been employed mainly in affinity chromatography [26][27][28][29][30][31][32][33][34][35][36][37][38][39]. Surprisingly, up to the present time, its adoption in other chromatographic modes has been rather scarce.…”

Section: Introductionmentioning

confidence: 99%

New multimodal stationary phases prepared by Ugi multicomponent approach

Sýkora

Záruba

Butnariu

et al. 2020

J of Separation Science

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Eight different stationary phases based on two aminopropyl silicas of different brands suitable for multimodal chromatography applications have been prepared by a four-component Ugi reaction. The intention was to synthesize stationary phases significantly differing in their properties hereby demonstrating flexibility of the Ugi synthetic protocol. Diverse functional groups including a nonpolar long aliphatic chain, phenyl moiety, cholic acid scaffold, phenylboronic and monosaccharide units, charged betaine, and arginine moieties were immobilized on a silica surface. The novel sorbents were extensively characterized by elemental analysis, Raman spectroscopy, and chromatography. Considering the anchored chemical structures covalently bonded to the silica surface, reversed-phase, hydrophilic, and ion-exchange separation modes were expected. The chromatographic evaluation was performed directed to map the potential of the individual columns specifically in the mentioned chromatographic modes. The Ugi synthetic protocol has proven to be a simple, feasible, and versatile tool for the synthesis of sorbents of variable properties. The newly prepared stationary phases differed considerably in hydrophobicity and ion-exchange ability. A significant influence of the supporting aminopropyl silica on the final chromatographic behavior was observed. Finally, one practical example confirming applicability of the newly prepared sorbents was demonstrated in separation of cytarabine.

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