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

Abstract：

: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.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Du D,Ju H,Zhang X,Chen J,Cai J,Chen H

doi

10.1021/bi0507332

subject

Has Abstract

pub_date

2005-08-30 00:00:00

pages

11539-45

issue

34

eissn

0006-2960

issn

1520-4995

journal_volume

44

pub_type

杂志文章

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