Telomerase catalyzed fluorescent probes for sensitive protein profiling based on one-dimensional microfluidic beads array.
A nucleic acid-based signal amplified method for multiple proteins detection based on one-dimensional beads array using telomerase catalyzed fluorescent probes has been developed in this paper. The biotin labeled fluorescent probes were synthesized by telomerase in homogeneous solution. Approximately 360-480 fluorescein molecules were inserted in each probe. The limit of detection for p53 protein is 1.1 pM (S/N=3) and a 3 orders of linear dynamic range is obtained. The sensitivity is nearly two orders higher than the two-site "sandwich" immunoassay using the same platform. Using this method, cellular p53 protein contents of as few as 85 CNE2 cells per mul sample can be determined specifically. The expression changes of p53, c-myc and beta-actin in CNE2 cells were further examined as a function of anti-cancer drug treatment, and the results are consistent with our previous reports. Compared with immuno-polymerase chain reaction and immuno-rolling circle amplification, this method is simple, fast, cheap and suitable for multi-protein analysis. The multiplexed proteins profiling of cellular samples should facilitate the new opportunities to the fundamental research of tumor development and progression, especially to the low abundant tumor-associated proteins analysis.
Wen
Jianhui
J
State Key Laboratory of Chemo, Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Biomedical Engineering Center, Hunan University, Changsha 410082, China.
Yang
Xiaohai
X
Wang
Kemin
K
Tan
Weihong
W
Zuo
Xinbing
X
Zhang
He
H
D005456:Fluorescent Dyes / Q000737:chemistry*; D020869:Gene Expression Profiling / Q000379:methods*; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D008863:Microspheres; D040081:Protein Array Analysis / Q000379:methods*; D013050:Spectrometry, Fluorescence / Q000379:methods*
D005456:Fluorescent Dyes