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目的 :建立一种蛋白质甘露糖化修饰的方法。方法 :用离子交换层析和钴亲和层析法 ,纯化重组HER 2 /neu配体结合区2 (LBD2 )蛋白 ,并将纯化后的LBD2蛋白进行甘露糖化修饰。新合成的LBD2拟糖蛋白经质谱法检测后 ,用间苯二酚 硫酸法进一步鉴定。结果 :纯化后LBD2蛋白的纯度可达 90 %。新合成的LBD2拟糖蛋白在质谱图上呈现相应于甘露糖修饰后蛋白质分子量的预期峰形。经间苯二酚 硫酸法检测 ,结合于LBD2蛋白上的化学基团为糖分子。结论 :获得了LBD2拟糖蛋白 ,为开展甘露糖受体介导的抗原提呈及肿瘤疫苗的实验研究打下了基础
Objective: To establish a method of protein mannosylation. Methods: The recombinant HER2 / neu ligand binding domain 2 (LBD2) protein was purified by ion exchange chromatography and cobalt affinity chromatography, and the purified LBD2 protein was mannosylated. The newly synthesized LBD2 glycoprotein was detected by mass spectrometry and further identified by resorcinol sulfuric acid method. Results: The purity of LBD2 protein was up to 90% after purification. The newly synthesized LBD2 glycoprotein shows the expected peak shape on the mass spectrum corresponding to the molecular weight of the mannose-modified protein. By resorcinol sulfuric acid assay, the chemical group bound to the LBD2 protein is a sugar molecule. CONCLUSION: The LBD2 mimic glycoprotein was obtained and laid the foundation for the experimental study of mannose receptor-mediated antigen presentation and tumor vaccine