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Glycochips from Polyanionic Glycopolymers as Tools for Detecting Shiga Toxins
Author(s) -
Uzawa Hirotaka,
Ito Hiroki,
Neri Paola,
Mori Hiroshi,
Nishida Yoshihiro
Publication year - 2007
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200700439
Subject(s) - surface plasmon resonance , glycopolymer , chemistry , sugar , shiga toxin , chromatography , polymer , nanotechnology , biochemistry , materials science , escherichia coli , organic chemistry , nanoparticle , copolymer , gene
An alternating layer‐by‐layer adsorption methodology was applied to the assembly of glycochips by using synthetic polyanionic glycopolymers. Three glycochips carrying globobioside (Gb 2 ), β‐lactoside ( β ‐Lac), or α ‐ D ‐mannoside ( α ‐Man) residues were prepared, and used for the detection of Shiga toxins, Stx‐1 and Stx‐2, by using surface plasmon resonance (SPR). Using this method, we could confirm that both Stx‐1 and Stx‐2 show binding specificity for the Gb 2 glycochip as well as a weak affinity for the β ‐Lac glycochip. The affinity constants of these toxins depended strongly on the sugar content of the Gb 2 polymer used to prepare the glycochip. Greater affinity was observed for chips with a higher sugar content (up to 43 %) in the Gb 2 glycopolymer. The maximal affinity constants of Stx‐1 and Stx‐2 ( K a =10 8 –10 9 M −1 ) enabled highly sensitive and facile analysis (10 ng mL −1 , 30 min). When Gb 2 glycopolymers were used as competitors, Stx‐1 and Stx‐2 behaved differently from one another in terms of their SPR response; this allowed us to perform discriminative analysis between the two toxins.