本文選題：固相法　切入點(diǎn)：丙型肝炎疫苗多肽　出處：《西北大學(xué)》2007年碩士論文

【摘要】： 本文以工業(yè)化生產(chǎn)為目的，研究了用Fmoc固相法人工合成丙型肝炎疫苗多肽(以下簡稱疫苗多肽)的生產(chǎn)工藝，比較了Wang樹脂和二氯三苯甲基樹脂(以下簡稱二氯樹脂)在固相合成中的的優(yōu)勢和劣勢，以及粗產(chǎn)物肽和目標(biāo)肽的產(chǎn)率和純度；創(chuàng)造性的引入N~α-Dmb基團(tuán)，提高了困難氨基酸Val的連接率；優(yōu)化了脫保護(hù)的策略以及切割試劑的選擇；通過質(zhì)譜鑒定分析所得目標(biāo)多肽的分子量，用RP-HPLC進(jìn)行粗肽的分析和純化，系統(tǒng)完整的建立了整套合成純化工藝路線及相關(guān)的工藝參數(shù)，為后續(xù)的工業(yè)化生產(chǎn)奠定了堅(jiān)實(shí)的基礎(chǔ)。采用該合成純化工藝路線，以二氯樹脂為載體，合成的粗肽產(chǎn)率達(dá)到了90.76％，純化后的目標(biāo)肽產(chǎn)率為63.02％純度達(dá)到了98.5％，并實(shí)現(xiàn)了18克每批的合成量。二氯樹脂以其高效率、低成本、產(chǎn)品的高純度完全符合工業(yè)化生產(chǎn)疫苗多肽的各項(xiàng)經(jīng)濟(jì)技術(shù)指標(biāo)。 論文包括以下四個(gè)部分： 1文獻(xiàn)綜述部分，簡述了丙型肝炎疫苗的研究現(xiàn)狀，詳細(xì)介紹了多肽固相合成技術(shù)中的合成方法、分離純化與鑒定技術(shù)，最后概述了多肽類藥物的應(yīng)用情況與研究展望。 2采用Fmoc固相法人工合成了疫苗多肽，比較了兩種樹脂在合成過程中的異同點(diǎn)，，對合成過程中的幾個(gè)重要步驟和難點(diǎn)進(jìn)行了優(yōu)化和改進(jìn)，如針對困難氨基酸Val的連接，設(shè)計(jì)了引入N~α-Dmb基團(tuán)的獨(dú)特合成策略，提高了連接率及切割效率，確定了合成工藝中相關(guān)的各項(xiàng)工藝技術(shù)參數(shù)。最后用二氯樹脂做為載體進(jìn)行了18克每批的疫苗多肽合成。 3用RP-HPLC進(jìn)行粗肽的分析和純化，根據(jù)疫苗多肽的理化性質(zhì)采用乙腈—水的流動(dòng)相體系，并確定了RP-HPLC分析和純化的條件。 4總結(jié)了實(shí)驗(yàn)結(jié)果，確定了疫苗多肽合成、純化工藝路線及相關(guān)的工藝技術(shù)參數(shù)。
[Abstract]:In this paper, the production process of hepatitis C vaccine polypeptide (hereinafter referred to as hepatitis C vaccine polypeptide) was studied by using Fmoc solid phase method for the purpose of industrial production. The advantages and disadvantages of Wang resin and dichlorotriphenylmethyl resin (hereinafter referred to as dichloromethyl resin) in solid phase synthesis, the yield and purity of crude product peptide and target peptide were compared, and the N- 偽 -Dmb group was introduced creatively. The binding rate of difficult amino acid Val was improved, the deprotection strategy and the selection of cutting reagent were optimized, the molecular weight of the target peptide was identified by mass spectrometry, and the crude peptide was analyzed and purified by RP-HPLC. A complete set of synthetic and purification process routes and related process parameters were established, which laid a solid foundation for subsequent industrial production. The synthesis and purification process route was adopted, and dichlororesin was used as carrier. The crude peptide yield reached 90.76%, the purity of the purified peptide was 63.02%, and the yield of 18 g / batch of dichlororesin was achieved by its high efficiency and low cost. The high purity of the product fully accords with the economic and technical indexes of the industrialized production of vaccine peptides. The thesis includes the following four parts:. 1 in the part of literature review, the research status of hepatitis C vaccine was briefly introduced, the synthesis methods, separation, purification and identification of peptide solid phase synthesis technology were introduced in detail. Finally, the application and prospect of polypeptide drugs were summarized. The peptide of vaccine was synthesized by Fmoc solid phase method. The similarities and differences between the two resins in the process of synthesis were compared. Several important steps and difficulties in the process of synthesis were optimized and improved, such as the connection of difficult amino acid Val. The unique synthesis strategy of introducing Na--Dmb group was designed, the connection rate and cutting efficiency were improved, and the relevant technological parameters were determined. Finally, 18 grams of vaccine polypeptides were synthesized with dichlororesin as carrier. 3The crude peptides were analyzed and purified by RP-HPLC. The mobile phase system of acetonitrile-water was used according to the physicochemical properties of vaccine peptides, and the conditions for RP-HPLC analysis and purification were determined. 4 the experimental results were summarized, and the synthesis, purification process and related technological parameters of vaccine polypeptide were determined.
【學(xué)位授予單位】：西北大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2007
【分類號】：R392

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 陰新強(qiáng);冉利;劉文虎;晏紀(jì)成;;大規(guī)模固相合成肽類藥物研究進(jìn)展[J];中國現(xiàn)代醫(yī)藥雜志;2010年12期

相關(guān)碩士學(xué)位論文 前1條

1 程朋真;免疫九肽質(zhì)量檢測方法的建立與穩(wěn)定性研究[D];西北大學(xué);2010年

本文編號：1678576