title: Energy transfer-based multiplexed assay of proteases by using gold nanoparticle and quantum dot conjugates on a surface.
abstract: Rapid and sensitive assay of proteases and their inhibition in a high-throughput manner is of great significance in the diagnostic and pharmaceutical fields. We developed a multiplexed assay system of proteases and their inhibition by measuring the energy transfer between quantum dots (QDs) and gold nanoparticles (AuNPs) on a glass slide. In this system, while the photoluminescence (PL) of donor QDs immobilized on a surface was quenched due to the presence of AuNPs (energy acceptor) in close proximity, the protease activity caused modulation in the efficiency of the energy transfer between the acceptor and donor, thus enabling the protease assay. In comparison to the QD-dye system, the conjugate of the QD-AuNP gave rise to higher energy transfer efficiency, resulting in quantitative assay of proteases with more sensitivity. When matrix metalloproteinase, caspase, and thrombin were tested, a multiplexed assay was successfully achieved since the AuNP could be used as a common energy acceptor in conjunction with QDs having different colors. Our system is anticipated to find applications in the diagnosis of protease-related diseases and screening of potential drugs with high sensitivity in a high-throughput way.
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Republic of Korea.
authors.forename: Young-Pil
authors.initials: YP
authors.lastname: Kim
authors.forename: Young-Hee
authors.initials: YH
authors.lastname: Oh
authors.forename: Eunkeu
authors.initials: E
authors.lastname: Oh
authors.forename: Sungho
authors.initials: S
authors.lastname: Ko
authors.forename: Min-Kyu
authors.initials: MK
authors.lastname: Han
authors.forename: Hak-Sung
authors.initials: HS
authors.lastname: Kim
mesh_terms: D004735:Energy Transfer; D005898:Glass; D006046:Gold / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry*; D010447:Peptide Hydrolases / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D045663:Quantum Dots; D012996:Solutions; D013050:Spectrometry, Fluorescence; D013499:Surface Properties
chemical_list: D012996:Solutions; D006046:Gold; D010447:Peptide Hydrolases