title: Globotriose-functionalized gold nanoparticles as multivalent probes for Shiga-like toxin.
abstract: Compared to monovalent carbohydrates, multivalent carbohydrate ligands exhibit significantly enhanced binding affinities to their interacting proteins. Here, we report globotriose (P(k) ligand)-functionalized gold nanoparticle (AuNP) probes for the investigation of multivalent interactions with the B(5) subunit of Shiga-like toxin I (B-Slt). Six P(k)-ligand-encapsulated AuNPs (P(k)-AuNPs) of varying particle size and linker length were synthesized and evaluated for their potential as multivalent affinity probes by using a surface plasmon resonance competition assay. The affinity of these probes for the interacting proteins was greatly affected by nanoparticle size, linker length, and ligand density on nanoparticle surface. For example, the 20-nm 20-P(k)-l-AuNP, which had a relatively long linker showed a >10(8)-fold increase in affinity compared with the mono P(k) ligand. This intrinsic high-affinity AuNP probe specifically captured the recombinant B-Slt from Escherichia coli lysate, and the resulting purity of the B-Slt was >95 %. We also developed a robust P(k)-AuNP-based detection method for Slt-I by combining the technique with silver enhancement.
authors.affiliation: Institute of Chemistry, and Genomics Research Centre, Academia Sinica, 128 Taipei, Taiwan.
authors.forename: Yuh-Yih
authors.initials: YY
authors.lastname: Chien
authors.forename: Mi-Dan
authors.initials: MD
authors.lastname: Jan
authors.forename: Avijit Kumar
authors.initials: AK
authors.lastname: Adak
authors.forename: Hsiao-Chien
authors.initials: HC
authors.lastname: Tzeng
authors.forename: Yen-Ping
authors.initials: YP
authors.lastname: Lin
authors.forename: Yu-Ju
authors.initials: YJ
authors.lastname: Chen
authors.forename: Ken-Tseng
authors.initials: KT
authors.lastname: Wang
authors.forename: Chien-Tien
authors.initials: CT
authors.lastname: Chen
authors.forename: Chia-Chun
authors.initials: CC
authors.lastname: Chen
authors.forename: Chun-Cheng
authors.initials: CC
authors.lastname: Lin
mesh_terms: D001419:Bacteria / Q000166:cytology; D015374:Biosensing Techniques / Q000295:instrumentation*; D006001:Glycoconjugates / Q000737:chemistry; D006046:Gold / Q000737:chemistry*; D008024:Ligands; D053768:Metal Nanoparticles / Q000032:analysis* / Q000737:chemistry*; D011485:Protein Binding; D022622:Shiga Toxin 1 / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012995:Solubility; D020349:Surface Plasmon Resonance; D014281:Trihexosylceramides / Q000138:chemical synthesis; D014312:Trisaccharides / Q000737:chemistry* / Q000378:metabolism; D014867:Water / Q000737:chemistry
chemical_list: D006001:Glycoconjugates; D008024:Ligands; D022622:Shiga Toxin 1; D014281:Trihexosylceramides; D014312:Trisaccharides; D014867:Water; C113460:globotriose; C018549:globotriaosylceramide; D006046:Gold