Title of article :
Improvement of proteome coverage using hydrophobic monolithic columns in shotgun proteome analysis
Author/Authors :
Wang، نويسنده , , Fangjun and Dong، نويسنده , , Jing and Ye، نويسنده , , Mingliang and Wu، نويسنده , , Ren’an and Zou، نويسنده , , Hanfa، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2009
Pages :
8
From page :
3887
To page :
3894
Abstract :
Monolithic columns are widely used in shotgun proteome analysis. However, it is difficult to increase the separation capability and proteome coverage by using conventionally organic polymer-based monolithic column due to the difficulty of controlling homogeneity of the overall pore structure (both pores and microglobules), which leads to relatively low column efficiency. Therefore, we studied the effect of constitute and percentage of porogenic solvent, functional monomer, column length, and separation gradient on the peak capacity and proteome coverage by methacrylate-based reversed phase monolithic columns. It was demonstrated that the porous property of the hydrophobic monolith, which was mainly determined by the porogenic solvent, was crucial to the proteome coverage when similar methacrylate monomer was utilized and a ternary porogenic solvent was adopted to prepare C12 monolithic column with relatively homogeneous overall pore structure. It was also shown that high proteome coverage could be reliably obtained with online multidimensional separation using totally monolithic columns system with the length of analytical column at 85 cm and reversed phase separation gradient at 210 min.
Keywords :
methacrylate , Porogenic solvent , Monolithic column , Proteome coverage , Shotgun proteome analysis , Online multidimensional separation
Journal title :
Journal of Chromatography A
Serial Year :
2009
Journal title :
Journal of Chromatography A
Record number :
1511973
Link To Document :
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