A multifunctional micro-fluidic system for dielectrophoretic concentration coupled with immuno-capture of low numbers of Listeria monocytogenes.
In this study, we demonstrated a micro-fluidic system with multiple functions, including concentration of bacteria using dielectrophoresis (DEP) and selective capture using antibody recognition, resulting in a high capture efficiency of bacterial cells. The device consisted of an array of oxide covered interdigitated electrodes on a flat silicon substrate and a approximately 16 microm high and approximately 260 microm wide micro-channel within a PDMS cover. For selective capture of Listeria monocytogenes from the samples, the channel surface was functionalized with a biotinylated BSA-streptavidin-biotinylated monoclonal antibody sandwich structure. Positive DEP (at 20 V(pp) and 1 MHz) was used to concentrate bacterial cells from the fluid flow. DEP could collect approximately 90% of the cells in a continuous flow at a flow rate of 0.2 microl min(-1) into the micro-channel with concentration factors between 10(2)-10(3), in sample volumes of 5-20 microl. A high flow rate of 0.6 microl min(-1) reduced the DEP capture efficiency to approximately 65%. Positive DEP attracts cells to the edges of the electrodes where the field gradient is the highest. Cells concentrated by DEP were captured by the antibodies immobilized on the channel surface with efficiencies of 18 to 27% with bacterial cell numbers ranging from 10(1) to 10(3) cells. It was found that DEP operation in our experiments did not cause any irreversible damage to bacterial cells in terms of cell viability. In addition, increased antigen expression (antigens to C11E9 monoclonal antibody) on cell membranes was observed following the exposure to DEP.
Yang
Liju
L
Birck Nanotechnology Center and Bindley Bioscience Center, Purdue University, West Lafayette, IN 47907, USA.
Banada
Padmapriya P
PP
Chatni
Mohammad R
MR
Seop Lim
Kwan
K
Bhunia
Arun K
AK
Ladisch
Michael
M
Bashir
Rashid
R
D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D000918:Antibody Specificity; D001710:Biotin / Q000737:chemistry; D004129:Dimethylpolysiloxanes; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods; D021902:Enterobacter aerogenes / Q000276:immunology / Q000302:isolation & purification; D013293:Enterococcus faecalis / Q000276:immunology / Q000302:isolation & purification; D008089:Listeria monocytogenes / Q000276:immunology / Q000302:isolation & purification*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D012825:Silicon / Q000737:chemistry; D012828:Silicones; D019809:Streptavidin / Q000737:chemistry
D000911:Antibodies, Monoclonal; D004129:Dimethylpolysiloxanes; D012828:Silicones; C013830:baysilon; D001710:Biotin; D019809:Streptavidin; D012825:Silicon