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Study on Synthesizing HCV Polypeptides by Solid-phase Method

Author: SunLiShu
Tutor: ChenWuLing
School: Northwestern University
Course: Microbiology
Keywords: Solid-phase method Hepatitis C vaccine polypeptide Polypeptide purification
CLC: R392
Type: Master's thesis
Year: 2007
Downloads: 245
Quote: 2
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Abstract


In this paper, for the purpose of industrial production , the solid phase method with Fmoc synthetic hepatitis C vaccine polypeptide (hereinafter referred to as the vaccine polypeptide ) Production compared Wang resin and dichloro - trityl resin (hereinafter referred to as dichloro resin ) in the solid-phase synthesis of the advantages and disadvantages, and the crude peptide and the target peptide , the yield and purity ; creative introduction of N ~ α-Dmb group, to improve the connection rate of the difficult amino acids Val ; optimized deprotection strategy as well as cutting the reagent of choice; identified by mass spectrometry analyzes of the molecular weight of the target polypeptide using RP-HPLC analysis and purification of crude peptide , a complete system to establish a set of synthetic purification process route and process parameters for subsequent industrial production laid a solid foundation . The process route for the synthesis purified to dichloro resin as a carrier , a synthetic crude peptide yield of 90.76% and 98.5% , after purification of the target peptide in a yield of 63.02% purity , and to achieve the synthesis of 18 grams of each batch amount . Dichloro resin for its high efficiency, low cost , high purity products fully comply with the economic and technical indicators for industrial production of vaccine peptides . The paper includes the following four parts : a literature review section briefly Hepatitis C Vaccine Research , details of the method for the synthesis of solid phase peptide synthesis technology , isolation , purification and identification techniques , and finally an overview of the application of peptides situation and research prospect . 2 by Fmoc solid-phase method and synthetic vaccines polypeptide , compare the similarities and differences of the two resins in the synthesis process , the optimization and improvement of several important steps in the synthesis process and difficult , as for the difficulties in amino acid Val connection of the design unique synthetic strategy for the introduction of the N ~ α-Dmb group connection rate and cutting efficiency , determine the parameters of the technology related synthesis process . Finally , as the carrier 18 g of each batch of vaccine peptide synthesis with methylene resin . 3 by RP-HPLC analysis and purification of the crude peptide , using a mobile phase of acetonitrile - water system the physicochemical properties of the polypeptide according to the vaccine , and to determine the conditions for the RP-HPLC analysis and purification . 4 summarizes the experimental results to determine the vaccine peptide synthesis and purification process route and related parameters of the process technology .

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CLC: > Medicine, health > Basic Medical > Medical Immunology
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