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中文摘要：
目的验证肺癌组织中热休克蛋白70（HSP 70）表达量,从肺癌组织中获得HSP 70并初步制备其人源性抗体,为进一步研究其生物功能和临床应用奠定基础。方法收集肺癌组织和癌旁组织标本,应用双向电泳-免疫印记方法验证HSP 70的表达量,经过亲和层析和离子交换层析方法获得肺癌组织中HSP 70作为抗原,经过四轮亲和-吸附-洗脱,从人源性肺癌噬菌体单链抗体库中初步筛选其抗体。结果双向电泳-免疫印记结果表明,肺癌组织HSP 70表达量高于癌旁组织。凝胶电泳和免疫印记结果表明,从肺癌组织中获得了人源性HSP 70,以此HSP 70作为抗原,初步筛选出了其抗体。结论人源性HSP 70及其人源性特异结合抗体的获得,为进一步研究其抗肺癌的临床应用提供了基础依据。
英文摘要：
Objective To validate the expression level of HSP 70 in the tissues of lung cancer, gain the human HSP 70 from lung cancer tissues and prepare its antibody, lay the foundation for further research about its biological function and clinical use. Methods Lung cancer tissues and cancerous lung cancer were collected, their expression level of HSP 70 were validated by using two - dimensional western blotting and it was separated by ConA- sepharose affinity column chromatography and EDTA negative charge column chromatography. HSP 70 extracted from lung cancer was used as antigen to biopan human scFv phage library of lung cancer through four rounds of biopanning. High affinity and specificity positive monoclonical phage antibody was identified by ELISA. Results Two - dimensional western blotting results showed the expression level of HSP 70 of lung cancer tissue was separated and purified. HSP 70 was used as antigen to biopan human antibody library, then its antibody was prepared. Conclusion The gaining of human HSP 70 and its human antibody lay the foundation for further research about its biological function and clinical use.
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