Globotriose-functionalized gold nanoparticles as multivalent probes for Shiga-like toxin.
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.
Chien
Yuh-Yih
YY
Institute of Chemistry, and Genomics Research Centre, Academia Sinica, 128 Taipei, Taiwan.
Jan
Mi-Dan
MD
Adak
Avijit Kumar
AK
Tzeng
Hsiao-Chien
HC
Lin
Yen-Ping
YP
Chen
Yu-Ju
YJ
Wang
Ken-Tseng
KT
Chen
Chien-Tien
CT
Chen
Chia-Chun
CC
Lin
Chun-Cheng
CC
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
D006001:Glycoconjugates; D008024:Ligands; D022622:Shiga Toxin 1; D014281:Trihexosylceramides; D014312:Trisaccharides; D014867:Water; C113460:globotriose; C018549:globotriaosylceramide; D006046:Gold