Quantitative intracellular molecular profiling using a one-dimensional flow system.
We report on the development of one-dimensional microfluidic bead arrays for rapid and quantitative molecular profiling of human cancer cells. This new bioanalytical platform integrates the rapid binding kinetics of suspension bead carriers, the multiplexing and encoding capabilities of gene/protein chips, and the liquid handling advantages of microfluidic devices. Using antibody-conjugated beads in a two-site "sandwich" format, we demonstrate that the proteomic contents of as few as 56 human lung epithelial cancer cells can be determined with high sensitivity and specificity. The results indicate that each cell contains approximately 6 x 10(5) copies of the tumor suppressor protein P53. We have further examined the expression changes of P53, c-Myc, and beta-Actin as a function of anticancer drug treatment and have validated these changes by using Western blotting. This ability to quantitatively analyze normal and diseased cells raises new possibilities in studying cancer heterogeneity and circulating tumor cells.
Zhou
Leiji
L
Biomedical Engineering Center, State Key Lab of Chemo/Biosensing & Chemometrics, College of Chemistry & Chemical Engineering, Hunan University, Changsha 410082, China.
Wang
Kemin
K
Tan
Weihong
W
Chen
Yunqing
Y
Zuo
Xinbing
X
Wen
Jianhui
J
Liu
Bin
B
Tang
Hongxing
H
He
Lifang
L
Yang
Xiaohai
X
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling / Q000295:instrumentation* / Q000379:methods*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods*; D011506:Proteins / Q000235:genetics / Q000378:metabolism
D011506:Proteins