A Novel Protocol to Entrap Active Urease in a Tetraethoxysilane-Derived Sol-Gel Thin-Film Architecture

Narang, Upvan; Prasad, Patil; Bright, Frank V.; Kumar, Arun; Kumar, N. D.; Malhotra, Bansi D.; Kamalasanan, M. N.; Chandra, Subhas

doi:10.1021/cm00046a004

Cited by 67 publications

(39 citation statements)

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“…Indeed, it was shown that enzymes retain their activity toward their target and antibodies are able to bind their corresponding antigen . These interesting properties triggered the development of biosensors for medical or environmental diagnosis using metalloproteins , enzymes , or antibodies entrapped in silica gels.…”

Section: Introductionmentioning

confidence: 99%

Aptamer entrapment in microfluidic channel using one‐step sol‐gel process, in view of the integration of a new selective extraction phase for lab‐on‐a‐chip

et al. 2017

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There is a great demand for integrating sample treatment into μTASs. In this context, we developed a new sol-gel phase for extraction of trace compounds in complex matrices. For this purpose, the incorporation of aptamers in silica-based gel within PDMS/glass microfluidic channels was performed for the first time by a one-step sol-gel process. The effective gel attachment onto microchannel walls and aptamer incorporation in the polymerized gel were evaluated using fluorescence microscopy. A good gel stability and aptamer incorporation inside the microchannel was demonstrated upon rinsing and over storage time. The ability of gel-encapsulated aptamers to interact with its specific target (either sulforhodamine B as model fluorescent target, or diclofenac, a pain killer drug) was assessed too. The binding capacity of entrapped aptamers was quantified (in the micromolar range) and the selectivity of the interaction was evidenced. Preservation of aptamers binding affinity to target molecules was therefore demonstrated. Dissociation constant of the aptamer-target complex and interaction selectivity were evaluated similar to those in bulk solution. This opens the way to new selective on-chip SPE techniques for sample pretreatment.

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Section: Introductionmentioning

confidence: 99%

Aptamer entrapment in microfluidic channel using one‐step sol‐gel process, in view of the integration of a new selective extraction phase for lab‐on‐a‐chip

et al. 2017

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“…These silicon oxide glasses are prepared by hydrolytic polymerization of silicon alkoxide precursors (Brinker and Scherer, 1990). A high chemical, mechanical, and thermal stability makes sol-gels ideal materials from an industrial perspective (Avnir et al, 1994) and the mild polymerization conditions render the process highly compatible with biomolecules (Das et al, 1998;Ellerby et al, 1992;Gill and Ballesteros, 1998;Narang et al, 1994). Reetz andco-workers (1996, 1997) recognized the advantages of these sol-gel materials for the immobilization of lipases.…”

Section: Introductionmentioning

confidence: 99%

Sol‐gel immobilization of serine proteases for application in organic solvents

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Reinhoudt

2001

Biotech & Bioengineering

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The serine proteases alpha-chymotrypsin, trypsin, and subtilisin Carlsberg were immobilized in a sol-gel matrix and the effects on the enzyme activity in organic media are evaluated. The percentage of immobilized enzyme is 90% in the case of alpha-chymotrypsin and the resulting specific enzyme activity in the transesterification of N-acetyl-L-phenylalanine ethyl ester with 1-propanol in cyclohexane is 43 times higher than that of a nonimmobilized lyophilized alpha-chymotrypsin. The activities of trypsin and subtilisin Carlsberg are enhanced with 437 and 31 times, respectively. The effect of immobilization on the enzyme activity is highest in hydrophobic solvents.

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“…An emerging method for immobilization of proteins is their entrapment into a porous, inorganic silicate matrix that is formed via a low-temperature sol-gel processing method. 10,11 Numerous reports have described both fundamental aspects of entrapped proteins, such as their conformation, 12-14 dynamics, 15-17 accessibility, 14 and stability, [31][32][33][34][35][36][37][38] and their many applications for catalysis, sensing, and affinity chromatography. 11 However, few reports describe the entrapment of kinases into sol-gel derived silica, 39 and no reports describe studies of entrapped protein tyrosine kinases.…”

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confidence: 99%

Entrapment of Src Protein Tyrosine Kinase in Sugar-Modified Silica

et al. 2004

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A novel sugar-modified silica has been used to entrap for the first time a protein tyrosine kinase (PTK). Silane precursors bearing covalently attached gluconamide moieties were used in combination with the biocompatible precursor diglycerylsilane (DGS) to generate sol-gel derived silica that was able to encapsulate highly active Src PTK and preserve the activity of the enzyme over multiple uses. The relative activity of the enzyme was assayed using a LANCE based fluorescence resonance energy transfer method involving time-gated detection of fluorescence from a europium labeled antiphosphotyrosine antibody and Cy5 labeled streptavidin upon mutual binding to biotinylated phosphopeptides. Using this detection method, with the antibody and streptavidin external to the sol-gel matrix, it was possible to detect the phosphorylation of peptides with molecular weights of up to 2300 Da using the entrapped enzyme in N-(3-triethoxysilylpropyl)gluconamide (GLTES) doped glasses. Src kinase-doped glasses, derived from precursors such as tetramethyl orthosilicate, tetraethyl orthosilicate, or DGS that did not contain GLTES, provided no detectable enzyme activity. The addition of 1 mM ATP to the GLTES/DGS sol before the encapsulation of the protein increased the activity of the enzyme in the resulting gel, likely through a ligand-based stabilization mechanism. The use of such a system for determination of PTK activity and inhibition is demonstrated, setting the stage for the development of chromatographic and microarray based methods for the screening of kinase inhibitors.

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A Novel Protocol to Entrap Active Urease in a Tetraethoxysilane-Derived Sol-Gel Thin-Film Architecture

Cited by 67 publications

References 1 publication

Aptamer entrapment in microfluidic channel using one‐step sol‐gel process, in view of the integration of a new selective extraction phase for lab‐on‐a‐chip

Aptamer entrapment in microfluidic channel using one‐step sol‐gel process, in view of the integration of a new selective extraction phase for lab‐on‐a‐chip

Sol‐gel immobilization of serine proteases for application in organic solvents

Entrapment of Src Protein Tyrosine Kinase in Sugar-Modified Silica

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