Reactive Semiconductor Nanocrystals for Chemoselective Biolabeling and Multiplexed Analysis
| Autor: | Kelly Boeneman, James B. Delehanty, Eunkeu Oh, Duane E. Prasuhn, Robert C. Triulzi, W. Russ Algar, Sara G. Becker-Catania, Igor L. Medintz, Guoliang Tao, Bradley Scott, Travis L. Jennings, Kim E. Sapsford, Vaibhav Jain, Samantha Spindel |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Materials science
Cell Survival Color General Physics and Astronomy Nanotechnology Multiplexing Substrate Specificity Enterotoxins chemistry.chemical_compound Semiconductor quantum dots Cell Line Tumor Quantum Dots Animals Humans Semiconductor nanocrystals General Materials Science Sulfhydryl Compounds Amines Maleimide Immunoassay Bioconjugation Staining and Labeling Cell Membrane General Engineering Biological Transport Flow Cytometry Immunohistochemistry Semiconductors chemistry Nanocrystal Quantum dot Amine gas treating |
| Zdroj: | ACS Nano. 5:5579-5593 |
| ISSN: | 1936-086X 1936-0851 |
| DOI: | 10.1021/nn201050g |
| Popis: | Effective biological application of nanocrystalline semiconductor quantum dots continues to be hampered by the lack of easily implemented and widely applicable labeling chemistries. Here, we introduce two new orthogonal nanocrystal bioconjugation chemistries that overcome many of the labeling issues associated with currently utilized approaches. These chemistries specifically target either (1) the ubiquitous amines found on proteins or (2) thiols present in either antibody hinge regions or recombinantly introduced into other proteins to facilitate site-specific labeling. The amine chemistry incorporates aniline-catalyzed hydrazone bond formation, while the sulfhydryl chemistry utilizes nanocrystals displaying surface activated maleimide groups. Both reactive chemistries are rapidly implemented, yielding purified nanocrystal-protein bioconjugates in as little as 3 h. Following initial characterization of the nanocrystal materials, the wide applicability and strong multiplexing potential of these chemistries are demonstrated in an array of applications including immunoassays, immunolabeling in both cellular and tissue samples, in vivo cellular uptake, and flow cytometry. Side-by-side comparison of the immunolabeled cells suggested a functional equivalence between results generated with the amine and thiol-labeled antibody-nanocrystal bioconjugates in that format. Three-color labeling was achieved in the cellular uptake format, with no significant toxicity observed while simultaneous five-color labeling of different epitopes was demonstrated for the immunolabeled tissue sample. Novel labeling applications are also facilitated by these chemistries, as highlighted by the ability to directly label cellular membranes in adherent cell cultures with the thiol-reactive chemistry. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|