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Amplification of flow‐microcalorimetry signal by means of multiple bioaffinity layering of lectin and glycoenzyme
Author(s) -
Gemeiner Peter,
Dočolomanský Peter,
Vikartovská Alica,
Štefuca Vladimír
Publication year - 1998
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1998.tb00525.x
Subject(s) - invertase , chemistry , isothermal microcalorimetry , immobilized enzyme , adsorption , concanavalin a , chromatography , agarose , biosensor , lectin , cellulose , glycidyl methacrylate , bead , enzyme , biochemistry , organic chemistry , materials science , polymer , quantum mechanics , in vitro , composite material , polymerization , enthalpy , physics
This paper demonstrates a positive influence of a special, stepwise technique of enzyme immobilization based on the biospecific adsorption of the glycoenzyme invertase on immobilized concanavalin A (Con A), subsequent adsorption of the free Con A on the immobilized invertase:Con A support and repeated adsorption of invertase on the support. A 3‐fold repetition of the same procedure designed preliminarily as bioaffinity layering afforded up to a 10‐fold increase in catalytic activity of the immobilized invertase. Reactive hydrogels based on bead cellulose and bead poly(glycidyl methacrylate) were used as immobilization supports for the preparation of these highly active preparations. The enhancement in catalytic activity of immobilized invertase preparations was demonstrated thermometrically, by flow microcalorimetry. Further attractive aspects for utilizing the signal amplification of biosensors with immobilized enzymes are discussed.