Multiplex detection of protease activity with quantum dot nanosensors prepared by intein-mediated specific bioconjugation. Anal Chem 2008 Nov 15;80(22):8649-55
Date
10/17/2008Pubmed ID
18922019Pubmed Central ID
PMC2677517DOI
10.1021/ac801562fScopus ID
2-s2.0-56449087616 (requires institutional sign-in at Scopus site) 164 CitationsAbstract
We report here a protease sensing nanoplatform based on semiconductor nanocrystals or quantum dots (QDs) and bioluminescence resonance energy transfer (QD-BRET) to detect the protease activity in complex biological samples. These nanosensors consist of bioluminescent proteins as the BRET donor, quantum dots as the BRET acceptor, and protease substrates sandwiched between the two as a sensing group. An intein-mediated conjugation strategy was developed for site-specific conjugation of proteins to QDs in preparing these QD nanosensors. In this traceless ligation, the intein itself is spliced out and excluded from the final conjugation product. With this method, we have synthesized a series of QD nanosensors for highly sensitive detection of an important class of protease matrix metalloproteinase (MMP) activity. We demonstrated that these nanosensors can detect the MMP activity in buffers and in mouse serum with the sensitivity to a few nanograms per milliliter and secreted proteases by tumor cells. The suitability of these nanosensors for a multiplex protease assay has also been shown.
Author List
Xia Z, Xing Y, So MK, Koh AL, Sinclair R, Rao JAuthor
Yun Xing Biostatistician I in the Surgery department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsBiosensing Techniques
Buffers
Cell Line, Tumor
Energy Transfer
Humans
Inteins
Luciferases, Renilla
Luminescence
Matrix Metalloproteinase 2
Matrix Metalloproteinase 7
Mice
Peptide Hydrolases
Quantum Dots
Sensitivity and Specificity
Substrate Specificity
Urokinase-Type Plasminogen Activator
