Integrated polymer chip for two-dimensional capillary gel electrophoresis.
Two-dimensional (2D) gel electrophoresis (GE) is one of the most powerful methods for nucleic acid and protein separation, but generally suffers from high laboratory efforts connected with high analysis costs. Therefore, we herein present the development of a miniaturized 2D capillary GE (CGE) device which allows for an efficient protein separation in analysis times of about 1.5 h. This integrated 2D-CGE chip comprises a first channel for isoelectric focussing (IEF), a second specially designed transfer channel, 300 parallel micro channels, each having a cross section of 50 microm x 50 microm, and buffer reservoirs. The present work discusses fabrication aspects, in particular the combination of different microfabrication technologies, experimental separation performances of isoelectric focussing (IEF) and CGE, and presents computer simulations and first experimental results of protein transfer from the first to the second dimension.
Griebel
Anja
A
Fluidics & Simulation Department, Institut für Mikrotechnik Mainz GmbH - IMM, Carl-Zeiss-Str. 18-20, 55129 Mainz, Germany.
Rund
Sabine
S
Schönfeld
Friedhelm
F
Dörner
Wolfgang
W
Konrad
Renate
R
Hardt
Steffen
S
D000818:Animals; D003198:Computer Simulation; D004566:Electrodes; D019075:Electrophoresis, Capillary / Q000191:economics / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004867:Equipment Design; D005936:Glucans; D006834:Hydrazines; D007525:Isoelectric Focusing; D009211:Myoglobin / Q000032:analysis; D011108:Polymers / Q000737:chemistry*
C417561:Alexa 488 hydrazide; D005936:Glucans; D006834:Hydrazines; D009211:Myoglobin; D011108:Polymers; C009109:pullulan