Isolation And Analyse Of Tumor Antigen And IgY

Immune system recognized tumor antigens are the molecular basis for cancer immunotheraphy. Along with the new understanding of tumor antigen,the development of tumor vaccine has got into a new stage.The indentification methods of tumor antigens have been developed in last decade ,providing a larger number of candidate antigens for clinical trials. In this thesis,the classification of tumor antigens and the identification methods of tumor antigens .meanwhile, the structure of LC-ESI-MS/MS was introduced and the indentification method of tumor antigens by LC-ESI-MS/MS was discussed. The experiment employed mild acid elution to eliminate tumor antigens from culture cell lines.Got rid of salt and condensed,The extration was sequenced and indentified by LC-ESI-MS/MS, and then the aminophenol constitutes of tumor antigens were reckoned ,which corresponding the mass number showed by mass spectrum. On the other hand , the pH of acid elution and the time of acid wash was studied and the effect of getting rid of salt between nanofiltratin and C18 was compared. Meanwhile, the regulation of double separation by HPLC was discussed. ImmunoglobulinY(IgY) was isolated and purified from yolk of the eggs using a two-step salt precipitation and one-step gelfiltration protocal, and the purification of IgY was analysed by SDS-PAGE. During the experiment,the variety of electrophonesis was studied ,then the best condition was selected. The figure of electrophonesis showed that the purification of IgY can reach 98%. It was concluded that the method of purification and isolation is effective and simple.