Stable and fluid ethylphosphocholine membranes in a poly(dimethylsiloxane) microsensor for toxin detection in flooded waters

被引：28

作者：

Phillips, KS ^{[1
]}

Dong, Y ^{[1
]}

Carter, D ^{[1
]}

Cheng, Q ^{[1
]}

机构：

[1] Univ Calif Riverside, Dept Chem, Riverside, CA 92521 USA

来源：

ANALYTICAL CHEMISTRY | 2005年 / 77卷 / 09期

关键词：

D O I：

10.1021/ac0500481

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Highly stable and fluid supported bilayer membranes were fabricated by fusion of positively charged ethylphosphocholine (DOPC+) vesicles into poly(dimethylsiloxane) (PDMS) microchannels for immunosensing of cholera toxin (CT) in flooded waters. Compared to phosphatidylcholine (PC) layers in the microchannels, DOPC+ membranes show exceptionally strong resistance to air-dry damage, as demonstrated by fluorescence recovery after photobleaching (FRAP) measurements and protein adsorption studies. In FRAP experiments, the mobile fraction of PC membranes was found to decrease by 10% upon drying/rehydration and the lateral diffusion coefficient decreased from 2.2 to 1.6 mu m(2)/s, whereas the mobile fraction and diffusion coefficient for DOPC+ membranes remain virtually unchanged during this process. Characterization by confocal microscopy reveals that only 1% of the DOPC+ membrane in the microchannels was removed by the drying/rehydration process, as compared to 11% for PC. Protein adsorption trends indicate that the charge of DOPC+ membranes allows for tuning of solution conditions to enable the desired protein-membrane interaction to predominate at the interface. A flow-based immunoassay for bacterial toxin was developed with 5% GM1/DOPC+ membranes in PDMS channels, and a detection limit of 250 amol for CT was obtained from the calibration curves. The assay was successfully applied to detection of CT spiked in water samples from the Santa Ana River, with nearly identical response and sensitivity.

引用

页码：2960 / 2965

页数：6

共 45 条

[1] Supported membrane nanodevices [J].

Anrather, D ;

Smetazko, M ;

Saba, M ;

Alguel, Y ;

Schalkhammer, T .

JOURNAL OF NANOSCIENCE AND NANOTECHNOLOGY, 2004, 4 (1-2) :1-22

[2] SIMPLE METHOD FOR PREPARATION OF HOMOGENEOUS PHOSPHOLIPID VESICLES [J].

BARENHOLZ, Y ;

GIBBES, D ;

LITMAN, BJ ;

GOLL, J ;

THOMPSON, TE ;

CARLSON, FD .

BIOCHEMISTRY, 1977, 16 (12) :2806-2810

[3] Calcium transport by sarcoplasmic reticulum Ca-ATPase can be investigated on a solid-supported membrane [J].

Bartolommei, G ;

Buoninsegni, FT ;

Moncelli, MR .

BIOELECTROCHEMISTRY, 2004, 63 (1-2) :157-160

[4]

Buoninsegni FT, 2001, J INORG BIOCHEM, V86, P448

[5]

Cézanne L, 1999, BIOCHEMISTRY-US, V38, P2779, DOI 10.1021/bi9818568

[6] Incidence and molecular analysis of Vibrio cholerae associated with cholera outbreak subsequent to the super cyclone in Orissa, India [J].

Chhotray, GP ;

Pal, BB ;

Khuntia, HK ;

Chowdhury, NR ;

Chakraborty, S ;

Yamasaki, S ;

Ramamurthy, T ;

Takeda, Y ;

Bhattacharya, SK ;

Nair, GB .

EPIDEMIOLOGY AND INFECTION, 2002, 128 (02) :131-138

[7] Formation and spreading of lipid bilayers on planar glass supports [J].

Cremer, PS ;

Boxer, SG .

JOURNAL OF PHYSICAL CHEMISTRY B, 1999, 103 (13) :2554-2559

[8] Rapid prototyping of microfluidic systems in poly(dimethylsiloxane) [J].

Duffy, DC ;

McDonald, JC ;

Schueller, OJA ;

Whitesides, GM .

ANALYTICAL CHEMISTRY, 1998, 70 (23) :4974-4984

[9] Bivalent-ion-mediated vesicle adsorption and controlled supported phospholipid bilayer formation on molecular phosphate and sulfate layers on gold [J].

Ekeroth, J ;

Konradsson, P ;

Höök, F .

LANGMUIR, 2002, 18 (21) :7923-7929

[10] Ganglioside microarrays for toxin detection [J].

Fang, Y ;

Frutos, AG ;

Lahiri, J .

LANGMUIR, 2003, 19 (05) :1500-1505

← 1 2 3 4 5 →