title: Nanocapillaries for open tubular chromatographic separations of proteins in femtoliter to picoliter samples.
abstract: We have recently examined the potential of bare nanocapillaries for free solution DNA separations and demonstrated efficiencies exceeding 10(6) theoretical plates/m. In the present work, we demonstrate the use of bare and hydroxypropylcellulose (HPC) coated open tubular nanocapillaries for protein separations. Using 1.5 microm inner diameter (i.d.) capillary columns, hydrodynamically injecting femto- to picoliter volumes of fluorescent or fluorescent dye labeled protein samples, utilizing a pneumatically pressurized chamber containing 1.0 mM sodium tetraborate solution eluent (typically 200 psi) as the pump, and performing on-column detection using a simple laser-induced fluorescence detector, we demonstrate efficiencies of close to a million theoretical plates/m while generating single digit microliter volumes of waste for a complete chromatographic run. We achieve baseline resolution for a protein mixture consisting of transferrin, alpha-lactalbumin, insulin, and alpha-2-macroglobulin.
authors: Department of Chemistry and Biochemistry, The University of Oklahoma, Norman, Oklahoma 73019, USA.
authors: Xiayan
authors: X
authors: Wang
authors: Chang
authors: C
authors: Cheng
authors: Shili
authors: S
authors: Wang
authors: Meiping
authors: M
authors: Zhao
authors: Purnendu K
authors: PK
authors: Dasgupta
authors: Shaorong
authors: S
authors: Liu
mesh_terms: D002482:Cellulose / Q000031:analogs & derivatives; D002845:Chromatography / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D006801:Humans; D011506:Proteins / Q000032:analysis*; D018401:Sample Size
chemical_list: D011506:Proteins; D002482:Cellulose; C008079:hydroxypropylcellulose