Affinity purification and enzymatic cleavage of inter-alpha inhibitor proteins using antibody and elastase immobilized on CIM monolithic disks

Lim, Yow‐Pin; Josić, Djuro; Callanan, Helen; Brown, Jeanne; Hixson, Douglas C.

doi:10.1016/j.chroma.2004.11.006

Cited by 37 publications

(24 citation statements)

References 18 publications

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“…However, there are a significant amount of reports on application of GMA-EDMA monolithic materials for the preparation of flowthrough bioreactors based on different enzymes, e.g. immobilized invertase [24], polynucleotide phosphorylase [25], glucose oxidase [26], citrate lyase, malate dehydrogenase, isocitrate dehydrogenase, lactate dehydrogenase [27], papain [28], lignin peroxidase [29], deoxyribonuclease [30,31], elastase [32], chymotrypsin [33], beta-secretase [34], pronase [35], pectin lyase [36] glucuronan lyase [37], pepsin [38] and peptide-N-glycosidase F [39].…”

Section: Introductionmentioning

confidence: 99%

Monolithic bioreactors: Effect of chymotrypsin immobilization on its biocatalytic properties

Ponomareva

Kartuzova

Влах

et al. 2010

Journal of Chromatography B

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Section: Introductionmentioning

confidence: 99%

Monolithic bioreactors: Effect of chymotrypsin immobilization on its biocatalytic properties

Ponomareva

Kartuzova

Влах

et al. 2010

Journal of Chromatography B

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“…The major advantage of this single-step approach already shown in Fig. 1 These disks were also used by Lim et al [62] for immobilization of another proteolytic enzyme -elastase, and used for digestion of inter-a inhibitor protein. Although the complete digestion could not be achieved at flow rates 0.25-2.0 mL/min, the speed of the reaction makes the disk reactor a good tool for preparative isolation of cleaved fragments that are used later as an intermediate for the synthesis of overlapping peptide arrays.…”

Section: Direct Immobilization Via Epoxy Functionalitiesmentioning

confidence: 96%

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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“…The papers describing reactors obtained via immobilisation of trypsin on glycidyl methacrylate-ethylene dimethacrylate (GMA-EDMA) or silica monoliths and intended for protein digestion were presented in the current literature [2][3][4][5][6][7]. However, there are a considerable number of reports on implementation of monolithic materials for the preparation of flow-through reactors on the basis of different enzymes such as elastase [8] polynucleotide phosphorylase [9], betasecretase [10], citrate lyase, malate dehydrogenase, isocitrate dehydrogenase, lactate dehydrogenase [11], papain [12], lignin peroxidase [13], deoxyribonuclease [14,15], trypsin [16], chymotrypsin [17], invertase [18], pronase [19], pectin lyase [20] and glucose oxidase [21].…”

Section: Introductionmentioning

confidence: 98%

Synthesis of a monolithic, micro-immobilised enzyme reactor via click-chemistry

2012

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An immobilised enzyme reactor (IMER) in the form of capillary monolith was developed for a micro-liquid chromatography system. The plain monolith was obtained by in situ thermal copolymerisation of glycidyl methacrylate and ethylene dimethacrylate in a fused silica capillary (200 × 0.53 mm ID) by using n-propanol/1,4-butanediol as porogen. The enzyme, α-chymotrypsin (CT), was covalently attached onto the monolith via triazole ring formation by click-chemistry. For this purpose, the monolithic support was treated with sodium azide and reacted with the alkyne carrying enzyme derivative. CT was covalently linked to the monolith by triazole-ring formation. The activity behaviour of monolithic IMER was investigated in a micro-liquid chromatography system by using benzoyl-L-tyrosine ethyl ester (BTEE) as synthetic substrate. The effects of mobile-phase flow rate and substrate feed concentration on the final BTEE conversion were investigated under steady-state conditions. In the case of monolithic IMER, the final substrate conversion increased with increasing feed flow rate and increasing substrate feed concentration. Unusual behaviour was explained by the presence of convective diffusion in the macropores of monolith. The results indicated that the monolithic-capillary IMER proposed for micro-liquid chromatography had significant advantages with respect to particle-based conventional high-performance liquid chromatography-IMERs.

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Affinity purification and enzymatic cleavage of inter-alpha inhibitor proteins using antibody and elastase immobilized on CIM monolithic disks

Cited by 37 publications

References 18 publications

Monolithic bioreactors: Effect of chymotrypsin immobilization on its biocatalytic properties

Monolithic bioreactors: Effect of chymotrypsin immobilization on its biocatalytic properties

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

Synthesis of a monolithic, micro-immobilised enzyme reactor via click-chemistry

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