The Preliminary Study Of New Device For The Gel Protein's Electro-Transfer

After the genome mapping of many organisms include human being have finished, the keystone of life sciences in the post-genome time turned to the important component-proteomics. The most practical analysis method-Two-dimensional gel electrophoresis (2-DE) combined with protein identification by mass spectrometry (MS) is currently the workhorse for proteomics. A quite new method for combination is capillary electrophoresis transfer (CE-transfer), it makes a big break in the time-consume and applicability compared with other link-methods.Here, we set up a new device for gel protein's electro-transfer. And try to actualize the vertical electroextraction and electro-ejection of gel protein. There are two changes in the base of CE-transfer: 1. using the uncoated fused-silica capillary and electrophoresis buffer with additive cationic surfactant-cetyl trimithyl ammonium bromide (CTAB) to produce reversed electro-osmotic flow (r-EOF); 2. appending an electrode in the nether end of the capillary to actualize the gel protein's electro-transfer under the control of tri-electrodes. Followed the former work, we proved the changes were viable by improved experiments and did some tests about correlative factors. In the study of vertical protein's electro-transfer, we had validated the counteraction of gravity by r-EOF, the operability of tri-electrodes in the process of transfer and the feasibility of electro-transfer toward the protein in the gel respectively by the preliminary test and electro-transfer experiment followed off-line identification.Taking CE-UV-detection as the off-line identification to simulate mass spectrometry, we made some achievements in the preliminary study, through the detective sensibility was not good enough. The vertical CE-transfer greatly shorten the time for identification of gel protein; the design of tri-electrodes made the electroextraction and electro-ejection easily and controllable; the device for transfer provide a advantage to achieve automatization for its cheapness and making simply. In a word, the device gives a new selection of robotization for gel protein's electro-transfer and connection from 2-DE to MADLI-MS.