Enzyme immobilization on epoxy- and 1,1'-carbonyldiimidazole-activated methacrylate-based monoliths | Health & Environmental Research Online (HERO)

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Citation
Tags

HERO ID

7448664

Reference Type

Journal Article

Title

Enzyme immobilization on epoxy- and 1,1'-carbonyldiimidazole-activated methacrylate-based monoliths

Author(s)

Bencina, K; Podgornik, A; Strancar, A; Bencina, M

Year

2004

Is Peer Reviewed?

Yes

Journal

Journal of Separation Science
ISSN: 1615-9306
EISSN: 1615-9314

Volume

Issue

10-11

Page Numbers

811-818

Language

English

PMID

15354558

DOI

10.1002/jssc.200401800

Web of Science Id

WOS:000222856300007

Abstract

Monolithic Convective Interaction Media (CIM) have been activated with epoxide and imidazole carbamate functionalities and used as supports for covalent immobilization of protein A, deoxyribonuclease I, and trypsin. The efficiency of immobilization for these proteins was determined from the amount of bound IgG, degradation of DNA, and hydrolysis of Nalpha-benzoyl-L-arginine ethyl ester, respectively. The respective biological activities of trypsin and the binding capacity of protein A immobilized via imidazole carbamate groups were 11.45 and 2.25 times higher than those obtained for epoxide matrix while they were practically equal for deoxyribonuclease I. The kinetics of immobilization was studied in detail for trypsin under dynamic conditions and revealed that the enzyme immobilized via imidazole carbamate groups already reached its highest activity in 5 min. In contrast, a much longer time was required for immobilization via epoxy groups.

Keywords

1,1â²-Carbonyldiimidazole; Activated matrix; CIM; Enzyme; Epoxide; Immobilization; Monoliths; Protein; Amino acids; Chemical bonds; Epoxy resins; Hydrolysis; Proteins; Protein A; Trypsin; Enzyme immobilization; deoxyribonuclease I; DNA; enzyme; epoxide; imidazole derivative; immunoglobulin G; methacrylic acid; protein A; trypsin; article; chemical reaction kinetics; DNA degradation; enzyme activity; enzyme immobilization; high performance liquid chromatography; hydrolysis; nonhuman; priority journal; Animals; Arginine; Calibration; Cattle; Chromatography, High Pressure Liquid; Deoxyribonuclease I; DNA; Dose-Response Relationship, Drug; Enzymes, Immobilized; Epoxy Compounds; Hydrogen-Ion Concentration; Hydrolysis; Imidazoles; Immunoglobulin G; Kinetics; Ligands; Methacrylates; Models, Chemical; Pancreas; Staphylococcal Protein A; Time Factors; Trypsin