Advanced cleanup process of the free-flow microfluidic device for protein analysis.
The treatment of samples preparation is generally recognized as a bottleneck for the rapid analysis of protein because of the off-chip performance in many cases. In this study, we used the charge characteristics of protein to develop a simple and rapid electro-microfluidic desalting system as an effective means of cleaning up protein sample. When we loaded a urea-rich protein sample and a buffer solution into a free-flow zone electrophoresis (FFZE) chamber, the microfluidic device was able to separate the charged protein sample and the non-charged urea. With a 90 V electric field in the FFZE chamber, the removal efficiency of the urea was about 88% and the recovery of the protein was 78%. In addition, the desalted protein sample used in this device showed significant improvement with respect to the MALDI-TOF-MS spectrum signal of a fusion protein, which was fused to the gold-binding polypeptide with enhanced green fluorescent protein, as a model protein. The inflow of the purified fusion protein sample can be successfully immobilized on the gold surface and analyzed by confocal fluorescence microscopy and surface plasmon resonance for biotechnological sensors.
Huh
Yun Suk
YS
Department of Chemical and Biomolecular Engineering (BK21 program), Korea Advanced Institute of Science and Technology, Daejeon 305-701, Republic of Korea.
Park
Tae Jung
TJ
Yang
KwangSuk
K
Lee
Eun Zoo
EZ
Hong
Yeon Ki
YK
Lee
Sang Yup
SY
Kim
Do Hyun
DH
Hong
Won Hi
WH
D000818:Animals; D015374:Biosensing Techniques; D004586:Electrophoresis / Q000295:instrumentation / Q000379:methods; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D014508:Urea / Q000737:chemistry
D011506:Proteins; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D014508:Urea