Quantitative Intracellular Molecular Profiling Using a One-Dimensional Flow System

• Published in: Analytical chemistry (Washington) 2006-09, Vol.78 (17), p.6246-6251
• Main Authors: Zhou, Leiji, Wang, Kemin, Tan, Weihong, Chen, Yunqing, Zuo, Xinbing, Wen, Jianhui, Liu, Bin, Tang, Hongxing, He, Lifang, Yang, Xiaohai
• Format: Article
• Language: English
• Subjects: Analytical biochemistry: general aspects, technics, instrumentation; Analytical chemistry; Analytical, structural and metabolic biochemistry; Animals; Arrays; Biological and medical sciences; Cell Line; Cells; Cricetinae; Fundamental and applied biological sciences. Psychology; Gene Expression - drug effects; Gene Expression Profiling - instrumentation; Gene Expression Profiling - methods; Humans; Lung cancer; Oligonucleotide Array Sequence Analysis - instrumentation; Oligonucleotide Array Sequence Analysis - methods; Proteins; Proteins - genetics; Proteins - metabolism; Proteomics; Tumors
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac060598e

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 ∼6 × 105 copies of the tumor suppressor protein P53. We have further examined the expression changes of P53, c-Myc, and β-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.