Digital microfluidics for automated proteomic processing.
Clinical proteomics has emerged as an important new discipline, promising the discovery of biomarkers that will be useful for early diagnosis and prognosis of disease. While clinical proteomic methods vary widely, a common characteristic is the need for (i) extraction of proteins from extremely heterogeneous fluids (i.e. serum, whole blood, etc.) and (ii) extensive biochemical processing prior to analysis. Here, we report a new digital microfluidics (DMF) based method integrating several processing steps used in clinical proteomics. This includes protein extraction, resolubilization, reduction, alkylation and enzymatic digestion. Digital microfluidics is a microscale fluid-handling technique in which nanoliter-microliter sized droplets are manipulated on an open surface. Droplets are positioned on top of an array of electrodes that are coated by a dielectric layer - when an electrical potential is applied to the droplet, charges accumulate on either side of the dielectric. The charges serve as electrostatic handles that can be used to control droplet position, and by biasing a sequence of electrodes in series, droplets can be made to dispense, move, merge, mix, and split on the surface. Therefore, DMF is a natural fit for carrying rapid, sequential, multistep, miniaturized automated biochemical assays. This represents a significant advance over conventional methods (relying on manual pipetting or robots), and has the potential to be a useful new tool in clinical proteomics.
Jebrail
Mais J
MJ
Department of Chemistry, University of Toronto, Canada.
Luk
Vivienne N
VN
Shih
Steve C C
SC
Fobel
Ryan
R
Ng
Alphonsus H C
AH
Yang
Hao
H
Freire
Sergio L S
SL
Wheeler
Aaron R
AR
D000818:Animals; D057205:Automation, Laboratory / Q000295:instrumentation / Q000379:methods; D002417:Cattle; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D012710:Serum Albumin, Bovine / Q000032:analysis
D011506:Proteins; D012710:Serum Albumin, Bovine