Nanocapillaries for open tubular chromatographic separations of proteins in femtoliter to picoliter samples.
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.
Wang
Xiayan
X
Department of Chemistry and Biochemistry, The University of Oklahoma, Norman, Oklahoma 73019, USA.
Cheng
Chang
C
Wang
Shili
S
Zhao
Meiping
M
Dasgupta
Purnendu K
PK
Liu
Shaorong
S
D002482:Cellulose / Q000031:analogs & derivatives; D002845:Chromatography / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D006801:Humans; D011506:Proteins / Q000032:analysis*; D018401:Sample Size
D011506:Proteins; D002482:Cellulose; C008079:hydroxypropylcellulose