Abstract |
A new strategy for assessing cell surface carbohydrates and P-glycoprotein (P-gp) expression status and quantifying the cell numbers with an electrochemical immunoassay was designed. In order to construct the base of the cytosensor, a novel 3-D architecture was initially fabricated by combining nitrogen-doped carbon nanotubes, thionine, and gold nanoparticles via a simple layer-by-layer method. The formed architecture provided an effective matrix for concanavalin A (Con A) binding and made the immobilized Con A hold high stability and bioactivity. On the basis of the specific recognition of cell surface mannosyl groups to Con A, the Con A/3-D architecture interface showed a predominant capability for cell capture. With another coupled signal amplification based on a enzymatic catalytic reaction of HRP toward the oxidation of thionine by the H(2)O(2), which was induced by two-step immunoreactions, the proposed cytosensor showed an excellent analytical performance for the detection of HeLa cells ranging from 8.0 x 10(2) to 2.0 x 10(7) cells mL(-1) with a limit of detection of 500 cells mL(-1). Moreover, with the use of preblocking procedures, the mannosyl groups and P-gp on single HeLa cell could be further detected to be (4 +/- 2) x 10(10) molecules of mannose moieties and 8.47 x 10(6) molecules of P-gp. This strategy offers great promise for sensitive detection of cancer cells and cell surface receptors and thus may help improve cancer diagnosis and treatment.
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Authors | Jing-Jing Zhang, Fang-Fang Cheng, Ting-Ting Zheng, Jun-Jie Zhu |
Journal | Analytical chemistry
(Anal Chem)
Vol. 82
Issue 9
Pg. 3547-55
(May 01 2010)
ISSN: 1520-6882 [Electronic] United States |
PMID | 20369831
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- ATP Binding Cassette Transporter, Subfamily B, Member 1
- Carbohydrates
- Nanotubes, Carbon
- Phenothiazines
- Gold
- thionine
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Topics |
- ATP Binding Cassette Transporter, Subfamily B, Member 1
(analysis, chemistry)
- Biosensing Techniques
(methods)
- Carbohydrates
(analysis, chemistry)
- Cell Membrane
(chemistry)
- Electrochemistry
(instrumentation, methods)
- Gold
(chemistry)
- HeLa Cells
- Humans
- Immunoassay
(methods)
- Metal Nanoparticles
(chemistry)
- Nanotubes, Carbon
(chemistry)
- Phenothiazines
(chemistry)
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