Electrochemical immunoassay of membrane P-glycoprotein by immobilization of cells on gold nanoparticles modified on a methoxysilyl-terminated butyrylchitosan matrix

Dan Du; Huangxian Ju; Xueji Zhang; Jing Chen; Jie Cai; Hongyuan Chen

doi:10.1021/bi0507332

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Electrochemical immunoassay of membrane P-glycoprotein by immobilization of cells on gold nanoparticles modified on a methoxysilyl-terminated butyrylchitosan matrix

Journal article

Peer reviewed

Electrochemical immunoassay of membrane P-glycoprotein by immobilization of cells on gold nanoparticles modified on a methoxysilyl-terminated butyrylchitosan matrix

Dan Du, Huangxian Ju, Xueji Zhang, Jing Chen, Jie Cai and Hongyuan Chen

Biochemistry (Easton), Vol.44(34), pp.11539-11545

08/30/2005

DOI: https://doi.org/10.1021/bi0507332

Handle:

https://hdl.handle.net/2376/111814

PMID: 16114890

Abstract

Antibodies - metabolism

Antibodies - chemistry

Humans

ATP-Binding Cassette, Sub-Family B, Member 1 - metabolism

K562 Cells

Cells, Immobilized - metabolism

Microscopy, Atomic Force

ATP-Binding Cassette, Sub-Family B, Member 1 - chemistry

Cell Membrane - metabolism

Cell Membrane - ultrastructure

Electrochemistry - methods

Immunoassay

A strategy to detect P-glycoprotein (P-gp) on cell membrane and quantify the cell number using electrochemical immunoassay was developed by effective surface immunoreactions and immobilization of cells on a highly hydrophilic interface, which was constructed by adsorption of colloidal gold nanoparticles on a methoxysilyl-terminated (Mos) butyrylchitosan modified glassy carbon electrode (Au-CS/GCE). Atomic force microscopy studies proved that the nanoparticles adsorbed on Mos-butyrylchitosan were efficient in preventing the cell leakage and retaining the activity of immobilized living K562/ADM leukemic cells. The incubation with P-gp monoclonal antibody and then the secondary alkaline phosphatase (AP) conjugated antibody introduced AP onto the K562/ADM cell immobilized on Au-CS/GCE. The bound AP led to an amperometric response of 1-naphthyl phosphate. Under optimal conditions the response was proportional to the logarithm of cell concentration in the range from 5.0 x 10(4) to 1.0 x 10(7) cells mL(-)(1) with a detection limit of 1.0 x 10(4) cells mL(-)(1). The results were comparable to flow cytometric analysis of P-gp expression. This proposed method was practical, convenient, and significant in the clinic and cytobiology.

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Title: Electrochemical immunoassay of membrane P-glycoprotein by immobilization of cells on gold nanoparticles modified on a methoxysilyl-terminated butyrylchitosan matrix
Creators: Dan Du - Key Laboratory of Analytical Chemistry for Life Science (Education Ministry of China), Department of Chemistry, Nanjing University, Nanjing 210093, PR China
Huangxian Ju
Xueji Zhang
Jing Chen
Jie Cai
Hongyuan Chen
Publication Details: Biochemistry (Easton), Vol.44(34), pp.11539-11545
Academic Unit: School of Mechanical and Materials Engineering
Publisher: United States
Identifiers: 99900547590001842
Language: English
Resource Type: Journal article

Electrochemical immunoassay of membrane P-glycoprotein by immobilization of cells on gold nanoparticles modified on a methoxysilyl-terminated butyrylchitosan matrix

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