本文選題：日本血吸蟲 + 免疫脂質(zhì)體�。� 參考：《中南大學(xué)》2011年碩士論文

【摘要】：目的 制備針對日本血吸蟲的第三代吡喹酮長循環(huán)免疫脂質(zhì)體,為日本血吸蟲病的靶向性治療奠定工作基礎(chǔ)。 方法 1、大量表達并純化SIEA26-28 ku-scFv 大量培養(yǎng)帶有pET32a/SIEA26-28 ku-scFv質(zhì)粒的BL21(DE3)大腸桿菌,IPTG誘導(dǎo)SIEA26-28 ku-scFv表達,用Ni2+柱純化目的蛋白,將純化蛋白進行蛋白濃度測定、SDS-PAGE檢測、Western Blot鑒定。 2、制備吡喹酮長循環(huán)脂質(zhì)體 采用薄膜蒸發(fā)超聲分散法制備吡喹酮長循環(huán)脂質(zhì)體并測定其包封率。 3、制備攜帶單鏈抗體的長循環(huán)免疫脂質(zhì)體 采用新型異型雙功能交聯(lián)劑SPDP交聯(lián)抗體與脂質(zhì)體,制備第三代包裹吡喹酮的長循環(huán)免疫脂質(zhì)體,計算抗體偶聯(lián)率。 結(jié)果 1、大量表達SIEA26-28 ku-scFv,通過Ni2+螯合親和層析法提純單鏈抗體,通過Western Blot檢測證明單鏈抗體能特異性識別日本血吸蟲SIEA。 2、成功制備吡喹酮長循環(huán)脂質(zhì)體,計算其包封率為56.1%。 3、成功制備攜帶單鏈抗體與吡喹酮的第三代長循環(huán)免疫脂質(zhì)體,計算抗體偶連率為21.3%。 結(jié)論 成功提取和純化單鏈抗體,制備攜帶單鏈抗體和吡喹酮的第三代長循環(huán)免疫脂質(zhì)體,為下一步行靶向治療血吸蟲病奠定了工作基礎(chǔ)。
[Abstract]:PurposeTo prepare the third generation praziquantel long circulating immune liposomes for schistosomiasis japonicum and lay a foundation for the targeted treatment of schistosomiasis japonicum.Method1. Express and purify SIEA26-28 ku-scFv in large quantitiesThe expression of SIEA26-28 ku-scFv was induced by E. coli BL21 (DE3) with pET32a/SIEA26-28 ku-scFv plasmid. The purified protein was purified by Ni2 column. The protein concentration was determined by SDS-PAGE and identified by Western Blot.Preparation of long circulating liposomes of praziquantelThe long circulating liposomes of praziquantel were prepared by evaporation ultrasonic dispersion method and the encapsulation efficiency was measured.3. Preparation of long circulating immune liposomes carrying scFvThe third generation long circulating immune liposomes coated with praziquantel were prepared by using a new type of bifunctional crosslinking agent SPDP crosslinking antibody with liposome. The coupling rate of antibody was calculated.Result1. SIEA26-28 ku-scFv was expressed in large quantities and purified by Ni2 chelating affinity chromatography. The detection of Western Blot showed that scFv could recognize Schistosoma japonicum SIEA specifically.2. The long circulating liposomes of praziquantel were successfully prepared and the encapsulation efficiency was calculated to be 56.1.3. The third generation of long-cycle immune liposomes carrying single-chain antibody and praziquantel were successfully prepared, and the antibody coupling rate was calculated to be 21.3%.ConclusionThe third generation of long-circulating immune liposomes carrying single-chain antibodies and praziquantel were successfully extracted and purified, which laid a foundation for the further targeted treatment of schistosomiasis.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R392

【引證文獻】

相關(guān)期刊論文 前1條

1 金一;余傳信;;蛋白質(zhì)及多肽類藥物長效化研究進展[J];中國血吸蟲病防治雜志;2012年05期

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本文編號：1740345