Rapid protein concentration, efficient fluorescence labeling and purification on a micro/nanofluidics chip.
Fluorescence analysis has proved to be a powerful detection technique for achieving single molecule analysis. However, it usually requires the labeling of targets with bright fluorescent tags since most chemicals and biomolecules lack fluorescence. Conventional fluorescence labeling methods require a considerable quantity of biomolecule samples, long reaction times and extensive chromatographic purification procedures. Herein, a micro/nanofluidics device integrating a nanochannel in a microfluidics chip has been designed and fabricated, which achieves rapid protein concentration, fluorescence labeling, and efficient purification of product in a miniaturized and continuous manner. As a demonstration, labeling of the proteins bovine serum albumin (BSA) and IgG with fluorescein isothiocyanate (FITC) is presented. Compared to conventional methods, the present micro/nanofluidics device performs about 10(4)-10(6) times faster BSA labeling with 1.6 times higher yields due to the efficient nanoconfinement effect, improved mass, and heat transfer in the chip device. The results demonstrate that the present micro/nanofluidics device promises rapid and facile fluorescence labeling of small amount of reagents such as proteins, nucleic acids and other biomolecules with high efficiency.
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210093, China.
Chen
C
Wang
Jun
J
Ouyang
De-Kai
DK
Ye
Jing-Juan
JJ
Xu
Hong-Yuan
HY
Chen
Xing-Hua
XH
Xia
D000818:Animals; D002417:Cattle; D004285:Dogs; D004867:Equipment Design; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D007074:Immunoglobulin G / Q000737:chemistry* / Q000302:isolation & purification; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D012709:Serum Albumin / Q000737:chemistry / Q000302:isolation & purification; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000302:isolation & purification; D013194:Staining and Labeling / Q000295:instrumentation*
D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; D016650:Fluorescein-5-isothiocyanate