Separation of non-denatured proteins using semi-crosslinked polyacrylamide capillary gel electrophoresis

This work presents an approach to build a high-performance,low-viscous and replaceable separation matrix,semi-crosslinked polyacrylamide(semi-CPA)capillary gel electrophoresis.Non-denatured basic proteins,such as lysozyme,cytochrome C,ribonuclease A and trypsin were separated.The impacts of monomer and cross-linker concentrations on protein separation were studied,and the ability of dynamic capillary inner wall coating was demonstrated.The UV absorption interference by semi-CPA gel matrix was successfully overcome by apartial filling technique,which results in sensitivity 20 times higher than other protein separation method.The excellent separation ability,reproducibility and dynamic coating ability made semi-CPA an ideal separation media in both capillary electrophoresis and microfluidic chip separation scheme.