重组MUC1-MBP融合蛋白的纯化及其安全性和稳定性研究

发布时间：2018-06-03 00:06

本文选题：MUC1 + 肿瘤疫苗　；参考：《吉林大学》2010年硕士论文

【摘要】： MUC1是粘蛋白家族成员,广泛分布于肿瘤细胞表面,是肿瘤疫苗研究的有效靶点。目前,国外学者相继研制出许多以MUC1为靶点的肿瘤疫苗。本研究室在国内首次研制重组MUC1-MBP融合蛋白肿瘤疫苗,我们已经构建重组MUC1-MBP融合蛋白表达质粒,对表达的蛋白进行了初步纯化,并在MUC1-MBP疫苗的药效学研究方面取得了可喜的进展。但是目前的纯化方法得到的重组MUC1-MBP融合蛋白的纯度还较低,其毒理学和稳定性的研究也没有充分进行。本研究从这三个方面着手,对该蛋白疫苗进行进一步研究。研究通过IPTG诱导MUC1-MBP融合蛋白在大肠杆菌中表达;制备MBP特异性抗体免疫亲和层析柱,进一步纯化MUC1-MBP融合蛋白;通过豚鼠全身过敏试验和急性毒性试验检测MUC1-MBP融合蛋白疫苗的安全性;通过恒温稳定加速试验确定MUC1-MBP融合蛋白的稳定性;并通过添加单一稳定剂寻求优化的MUC1-MBP融合蛋白的保存条件。通过改进纯化工艺,重组MUC1-MBP融合蛋白的纯度明显提高;经安全性和稳定性的初步研究,结果表明重组MUC1-MBP融合蛋白安全、稳定,该结论为进一步研究开发MUC1-MBP疫苗奠定了基础。
[Abstract]:MUC1 is a member of mucin family, widely distributed on the surface of tumor cells, and is an effective target of tumor vaccine research. At present, many tumor vaccines targeting MUC1 have been developed by foreign scholars. The recombinant MUC1-MBP fusion protein tumor vaccine was developed in our laboratory for the first time in China. We have constructed the recombinant MUC1-MBP fusion protein expression plasmid, purified the expressed protein, and made gratifying progress in the pharmacodynamics of MUC1-MBP vaccine. However, the purity of recombinant MUC1-MBP fusion protein obtained by the present purification method is still low, and the toxicology and stability of the recombinant MUC1-MBP fusion protein have not been fully studied. In this study, the protein vaccine was further studied from these three aspects. The expression of MUC1-MBP fusion protein in Escherichia coli was induced by IPTG, and the MUC1-MBP fusion protein was further purified by preparing MBP specific antibody immunoaffinity chromatography column. The safety of MUC1-MBP fusion protein vaccine was detected by whole body allergy test and acute toxicity test, and the stability of MUC1-MBP fusion protein was determined by constant temperature stability accelerated test. The optimal preservation conditions of MUC1-MBP fusion protein were obtained by adding a single stabilizer. The purity of the recombinant MUC1-MBP fusion protein was improved by improving the purification process, and the safety and stability of the recombinant MUC1-MBP fusion protein were studied. The results showed that the recombinant MUC1-MBP fusion protein was safe and stable. This conclusion laid a foundation for further research and development of MUC1-MBP vaccine.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R392

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