A review on preparative and semi-preparative offgel electrophoresis for multidimensional protein/peptide assessment

作者：Moreda Pineiro Antonio^*; Garcia Otero Natalia; Bermejo Barrera Pilar

来源：Analytica Chimica Acta, 2014, 836: 1-17.

DOI：10.1016/j.aca.2014.04.053

• Offgel electrophoresis
• Isoelectric focusing
• Protein
• Peptide

Mass spectrometry (MS) techniques are commonly used for protein identification and further analysis of selected protein spots after high resolution 2-D electrophoresis. Complementary gel-free approaches have been developed during the last few years and have shown to be useful tools in modern proteomics. The development and application of various gel-free electrophoresis devices for performing protein fractionation according to the pI differences is therefore a topic of interest. This review describes the current state of isoelectric focusing (IEF) gel-free electrophoresis based on the Agilent offgel 3100 fractionator. The review includes, therefore, (i) an overview on IEF as well as other previous IEF gel-free electrophoresis developments; (ii) offgel fundamentals and future trends; (iii) advantages and disadvantages of current offgel procedures; (iv) requirements of isolated protein pellets for further offgel fractionation; (v) offgel fraction requirements to perform the second dimensional analysis by advance electrophoresis and chromatographic techniques; and (vi) effect of the offgel operating conditions on the stability of metal-protein complexes.

• 出版日期2014-7-11