Study On Synthesizing HCV Polypeptides By Solid-phase Method

Aiming at industrialized production, this paper focuses on the artificial technology of vaccine polypeptide of Hepatitis C with solid phase method. In the solid-phase synthesis, the advantages and disadvantages of Wang Resin and 2-chlorotrityl choloride Resin as well as the yield and purity of eribble product of peptide have been compared. N~α-Dmb group has been creatively inlet to elevate the connective rate of bary-amino acid Val and optimize the strategy of deprotection and the selection of cutting agent. The molecule weight of goal polypeptide has been evaluated and analyzed by MS. Eribble product of peptides have been analyzed and depurated by RP-HPLC. A complete set of schedule of synthsis and depuration and related parameters have been established systematically, which provides a substantial basis to the future production. In this process, the yield of errible products reached 90.76% and that of the final ones gets 63.02%, what is more, the fineness is 98.5% with Dichlorine Resin as the carrying agent. In addition, the amount of product has realized 18 gram per batch. 2-chlorotrityl choloride Resin can meet the requirements of all the economical specification because of its appropriate high efficiency, low cost and high purity.Four parts are included in this paper:1 Review . In this part, the current research and development of Hepatitis C vaccine is illustrated. The methods of synthesis, purification, separation and identification of solid-phase method are stated in detail. Finally, the application and research outlook of polypeptide medicine is summarized. 2 The vaccine peptide has been synthesized with Fmoc solid phase method. After comparing the two resins, some significant procedure and nodus have been improved. Such as the peculiar synthesis strategy of N~α-Dmb group inlet to aim directly at bary-amino acid Val, the elevation of connective rate and cutting rate as well as the determination of the parameters in this process. Thus, 18 gram per batch can be achieved with 2-chlorotrityl choloride Resin as the carrying agent.3 The rough peptide is analyzed and purified by RP-HPLC and the figure of the result has been compared for the purpose of optimizing the experiment condition.4 To summarize the experimental result and definite the parameters of synthesis and purification.