Microfluidic protein detection through genetically engineered bacterial cells.
Protein microarray technology, in which a large number of capture ligands are spatially arrayed at a high density, presents an attractive method for high-throughput proteomic analysis. Toward this end, we demonstrate the first cell-based protein detection in a microsystem, wherein Escherichia coli cells are genetically engineered to express the desired capture proteins on the membrane surface and are spatially arrayed as sensing elements in a microfluidic device. An E. coli clone expressing peptide ligands with high affinity and high specificity for target molecules was isolated a priori. Then these cells were electrokinetically immobilized on gold electrodes using dielectrophoresis, thus allowing each sensor element to be electrically addressable. Flow cytometry and subsequent fluorescence analysis verified the highly specific capture and detection of target molecules by the bacteria. Finally, through the coexpression of peptide-based capture ligands on the cell surface and fluorescent protein in the cytoplasm, we demonstrate an effective means of directly linking the fluorescence intensity to the density of capture ligands.
Oh
Sang-Hyun
SH
California Nanosystems Institute and Biomolecular Science and Engineering Program, University of California-Santa Barbara, California 93106, USA.
Lee
Sang-Ho
SH
Kenrick
Sophia A
SA
Daugherty
Patrick S
PS
Soh
Hyongsok T
HT
D004926:Escherichia coli / Q000235:genetics; D005434:Flow Cytometry; D005453:Fluorescence; D005818:Genetic Engineering / Q000379:methods; D008024:Ligands; D040081:Protein Array Analysis / Q000379:methods*; D011506:Proteins / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*
D008024:Ligands; D011506:Proteins