[Stabilization of polyacrylamide gel immobilized horseradish peroxidase by its covalent coupling to albumin].
Authors: Ugarova, Kershengol'ts, Artomonov, Berezin
Journal: Biokhimiia (Moscow, Russia)
Summary
# Editorial Summary Horseradish peroxidase (HRP) is widely used in diagnostic and research applications across equine medicine, but its thermal instability limits shelf-life and reliability in field conditions. Ugarova and colleagues investigated whether pre-coupling HRP to albumin via glutaraldehyde cross-linking before immobilisation in polyacrylamide gel (PAAG) could enhance enzyme stability and catalytic performance. The researchers systematically optimised gel composition and oligomer ratios, discovering that enzyme-albumin complexes at a 2.4:1 albumin-to-peroxidase ratio, immobilised in 40% PAAG, achieved maximal specific activity of 4.5 nmol/g whilst demonstrating 15-fold greater thermal stability than free enzyme and 250-fold greater stability than non-pretreated immobilised enzyme. This marked improvement in thermostability—achieved through albumin's protective effect on the peroxidase structure during immobilisation—suggests that enzyme-stabilisation techniques could substantially extend the practical utility of enzyme-based diagnostic assays in equine practice, particularly where temperature control during transport or storage is challenging.
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Practical Takeaways
- •This research is not applicable to equine practice; it is fundamental biochemistry research on enzyme stabilization techniques for laboratory use
Key Findings
- •Covalent coupling of horseradish peroxidase to albumin via glutaraldehyde increased thermostability and specific activity of polyacrylamide gel-immobilized enzyme
- •Optimal oligomers in 40% PAAG achieved maximal specific activity of 4.5 nmol/g
- •Albumin/peroxidase ratio of 2.4 in 40% PAAG provided 15-fold greater stability than soluble enzyme and 250-fold greater than untreated immobilized enzyme