Intrinsic fluorescent recognition ligand scaffold based on chaperonins and water-soluble semiconductor quantum dots

Hongzhi Xie; Basil I. Swanson; Hiromi K. Kawaga; Jonathan D. Trent; Mudalige Kumara; Thomas Ippolito; Mircea Cotlet

doi:10.1117/12.809576

16 February 2009 Intrinsic fluorescent recognition ligand scaffold based on chaperonins and water-soluble semiconductor quantum dots

Hongzhi Xie, Basil I. Swanson, Hiromi K. Kawaga, Jonathan D. Trent, Mudalige Kumara, Thomas Ippolito, Mircea Cotlet

Author Affiliations +

Proceedings Volume 7188, Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications VI; 718807 (2009) https://doi.org/10.1117/12.809576
Event: SPIE BiOS, 2009, San Jose, California, United States

Abstract

We describe genetic engineering of a novel protein-nanoparticle hybrid system with great potential for patterning of various types of nanoparticles and for biosensing applications. The hybrid system is based on a genetically-modified chaperonin protein from the hyperthermophilic archaeon Sulfolobus shibatae. This chaperonin is an 18-subunit double ring, which self-assembles in the presence of Mg ions and ATP. We describe a chaperonin mutant (His-β- loopless:HBLL), with increased access to the central cavity and His-tags on each subunit extending into the central cavity. This mutant binds water-soluble semiconductor quantum dots, creating a protein-encapsulated fluorescent nanoparticle. By adding selective binding sites to the solvent-exposed regions of the chaperonin, this proteinnanoparticle bioconjugate becomes a sensor for specific targets. Using a combination of biochemical and spectroscopic assays, we characterize the formation, stoichiometry, affinity and stability of these novel sensors.

Citation Download Citation

Hongzhi Xie, Basil I. Swanson, Hiromi K. Kawaga, Jonathan D. Trent, Mudalige Kumara, Thomas Ippolito, and Mircea Cotlet "Intrinsic fluorescent recognition ligand scaffold based on chaperonins and water-soluble semiconductor quantum dots", Proc. SPIE 7188, Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications VI, 718807 (16 February 2009); https://doi.org/10.1117/12.809576

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