title: Ion-permeable membrane for on-chip preconcentration and separation of cancer marker proteins.
abstract: Cancer marker proteins have been electrophoretically concentrated and then separated in a microfluidic device. On-chip preconcentration was achieved using an ion-permeable membrane, consisting of acrylamide, N,N'-methylene-bisacrylamide and 2-(acrylamido)-2-methylpropanesulfonate. This negatively charged membrane was photopolymerized in the microdevice near the injection intersection. Anionic proteins were excluded from the porous membrane based on both size and charge, which concentrated target components in the injection intersection prior to separation by microchip capillary electrophoresis (μ-CE). Bovine serum albumin was used in the initial characterization of the system and showed a 40-fold enrichment in the μ-CE peak with 4 min of preconcentration. Adjustment of buffer pH enabled baseline resolution of two cancer biomarkers, α-fetoprotein (AFP) and heat shock protein 90 (HSP90), while fine control over preconcentration time limited peak broadening. Our optimized preconcentration and μ-CE approach was applied to AFP and HSP90, where enrichment factors of >10-fold were achieved with just 1 min of preconcentration. Overall, the process was simple and rapid, providing a useful tool for improving detection in microscale systems.
authors.affiliation: Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602, USA.
authors.forename: Pamela N
authors.initials: PN
authors.lastname: Nge
authors.forename: Weichun
authors.initials: W
authors.lastname: Yang
authors.forename: Jayson V
authors.initials: JV
authors.lastname: Pagaduan
authors.forename: Adam T
authors.initials: AT
authors.lastname: Woolley
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000737:chemistry / Q000302:isolation & purification*; D002417:Cattle; D046209:Electrophoresis, Microchip / Q000295:instrumentation* / Q000379:methods; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008567:Membranes, Artificial; D009363:Neoplasm Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification; D000509:alpha-Fetoproteins / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D014408:Biomarkers, Tumor; D018841:HSP90 Heat-Shock Proteins; D008567:Membranes, Artificial; D009363:Neoplasm Proteins; D000509:alpha-Fetoproteins; D012710:Serum Albumin, Bovine