Development of a chromatographic bioreactor based on immobilized β-glucuronidase on monolithic support for the determination of dextromethorphan and dextrorphan in human urine

Calleri, Enrica; Marrubini, Giorgio; Massolini, Gabriella; Lubda, Dieter; Fazio, S.S. de; Furlanetto, Sandra; Wainer, Irving W.; Manzo, Luigi; Caccialanza, Gabriele

doi:10.1016/j.jpba.2004.02.033

Cited by 20 publications

(15 citation statements)

References 22 publications

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“…9e, this also begins by converting epoxy groups on a monolith like GMA/EDMA into diol groups. These diol groups are next reacted with DSC to place succinimidyl carbonate groups on the monolith's surface [4,36,75]; an amine-containing monolith can also be used for this reaction [57]. The activated form of the monolith is then reacted with a ligand such as a protein that con-…”

Section: Covalent Immobilization Methodsmentioning

confidence: 99%

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Affinity monolith chromatography

Mallik

Hage²

2006

J of Separation Science

190

282

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The combined use of monolithic supports with selective affinity ligands as stationary phases has recently given rise to a new method known as affinity monolith chromatography (AMC). This review will discuss the basic principles behind AMC and examine the types of supports and ligands that have been employed in this method. Approaches for placing affinity ligands in monoliths will be considered, including methods based on covalent immobilization, biospecific adsorption, entrapment, and the formation of coordination complexes. Several reported applications will then be presented, such as the use of AMC for bioaffinity chromatography, immunoaffinity chromatography, immobilized metal-ion affinity chromatography, dye-ligand affinity chromatography, and biomimetic chromatography. Other applications that will be discussed are chiral separations and studies of biological interactions based on AMC.

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Section: Covalent Immobilization Methodsmentioning

confidence: 99%

“…The DSC method has been used with GMA/ EDMA monoliths to immobilize HSA [4] and with an amine-containing silica monolith to immobilize b-glucuronidase [57]. This method is fast and has been reported to be complete within 10 h [4].…”

Section: Covalent Immobilization Methodsmentioning

confidence: 99%

Affinity monolith chromatography

Mallik

Hage²

2006

J of Separation Science

190

282

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“…Along with traditional silica particles, silica monoliths [82][83][84] and sol-gels have been used with affinity ligands [85,86]. Affinity ligands can be immobilized in silica monoliths using methods similar to those already described for silica particles.…”

Section: Other Immobilization Methodsmentioning

confidence: 99%

“…An immunoaffinity/protein A dual column system for the determination of HSA and IgG in serum samples [82]. Diagram (a) depicts the connections during sample application.…”

Section: Use Of Silica and Affinity Ligands For Direct Detectionmentioning

confidence: 99%

Applications of silica supports in affinity chromatography

Schiel

Mallik

Soman

et al. 2006

J of Separation Science

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The combined use of silica-based chromatographic supports with immobilized affinity ligands can be used in many preparative and analytical applications. One example is the use of silica-based affinity columns in HPLC, giving rise to a method known as high-performance affinity chromatography (HPAC). This review discusses the role that silica has played in the development of affinity chromatography and HPAC and the applications of silica in these methods. This includes a discussion of the types of ligands that have been employed with silica and the methods by which these ligands have been immobilized. Various formats have also been presented for the use of silica in affinity chromatographic methods, including assays involving direct or indirect analyte detection, on-line or off-line affinity extraction, and chiral separations. The use of silica-based affinity columns in studies of biological systems based on zonal elution and frontal analysis methods will also be considered.

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“…In continuing efforts, this group also immobilized b-glucuronidase on a silica monolith and used it in the same integrated system for determination of dextromethorphan and dextrorphan in urine [37]. This time they used monolith modified with 3-aminopropyltrimethoxysilane and activated with N,N'-disuccinimidyl carbonate.…”

Section: Activated Silica Columnmentioning

confidence: 99%

Less common applications of monoliths: I. Microscale protein mapping with proteolytic enzymes immobilized on monolithic supports

Švec

2006

Electrophoresis

131

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This review summarizes the recent contributions to the rapidly growing area of immobilized enzymes employing both silica and synthetic polymer-based monoliths as supports. Focus is mainly on immobilized proteolytic enzyme reactors designed for studies in proteomics. Porous monoliths emerged first as a new class of stationary phases for HPLC in the early 1990s. Soon thereafter, they were also used as supports for immobilization of proteins and preparation of both stationary phases for bioaffinity chromatography and enzymatic reactors. Organic polymer-based monoliths are typically prepared using a simple molding process carried out within the confines of a "mold" such as chromatographic column or capillary. Polymerization of a mixture comprising monomers, initiator, and porogenic solvent affords macroporous materials. In contrast, silica-based monoliths are first formed as a rigid rod from tetraalkoxysilane in the presence of PEG and subsequently encased with a plastic tube. Both types of monolith feature large through-pores that enable a rapid flow-through. Since all the solutions must flow through the monolith, the convection considerably accelerates mass transfer within the monolith. As a result, reactors including enzyme immobilized on monolithic support exhibit much higher activity compared to the reactions in solution.

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Development of a chromatographic bioreactor based on immobilized β-glucuronidase on monolithic support for the determination of dextromethorphan and dextrorphan in human urine

Cited by 20 publications

References 22 publications

Affinity monolith chromatography

Affinity monolith chromatography

Applications of silica supports in affinity chromatography

Less common applications of monoliths: I. Microscale protein mapping with proteolytic enzymes immobilized on monolithic supports

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