𝔖 Bobbio Scriptorium
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Electrosynthetic immobilisation of proteins for bioanalysis

✍ Scribed by D. J. Walton; C. J. Campbell; P. G. Richards; J. Heptinstall

Publisher
John Wiley and Sons
Year
1998
Tongue
English
Weight
117 KB
Volume
8
Category
Article
ISSN
1616-301X

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✦ Synopsis

Use of electrosynthetic methodology allows the production of hen egg-white lysozyme (HEWL) either mononitrated at tyrosine 23 or bisnitrated at tyrosines 20 and 23, but never nitrated at tyrosine 53. This is a different sequence from that obtained by the chemical nitrating agent tetranitromethane, and when reduced by dithionite, the selectively modified enzyme can be anchored at pH 5 via the unique aromatic amino group to magnetic beads or other suitable matrices. HEWL so immobilised loses less than 10% of cell-wall lytic activity compared with the approximately 50% loss of activity when immobilised by conventional methodology at pH 9 via essentially random reaction at lysine residues and other functionalities which are nucleophilic at this pH. This result offers promise as a general method for selective protein immobilisation in biosensors and similar applications. *

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