This paper demonstrates the use of UV-transparent replaceable polymer networks for the separation of SDS-protein complexes on the basis of molecular weight. First, the use of linear (i.e. non-cross-linked) polyacrylamide is shown to provide molecular separation of SDS—protein complexes. A study reveals such columns to yield significantly greater lifetime than crosslinked gels because of the flexibility of the noncovalently attached polymer chains. However, column lifetime was still found to be limited ( ~ 20-40 injections), and detection at 214 nm was problematical because of the absorbance of polyacrylamide. UV-transparent polymer networks of dextran and PEG were substituted for polyacrylamide with successful molecular weight sieving of SDS—protein complexes at 214 nm. Due to their low to moderate viscosities, these networks could be routinely replaced leading to the possbillty of hundreds of lnjections with a single column. Migration time reproducibilities of 0.5 % RSD or less were found with replacement of the network. Using dextran, calibration plots of peak area vs concentration of standard protein were linear over the range of 0.5 µg/mL up to at least 0.25 mg/mL. Furthermore, plasma samples could be directly utilized because of the strong solvating power of SDS. Rapid separation of protein mixtures are demonstrated with these UV-transparent polymer networks.
ASJC Scopus subject areas
- Analytical Chemistry