| 摘要: |
| ［摘要］ 目的 研究应用抗体基因芯片检测鼻咽癌组织特异性蛋白质表达谱的新技术方案。方法 利用噬菌体抗体库筛选出抗鼻咽癌的特异性抗体库，以扩增抗体基因V-D-J片段作为标识分子用于制备抗体基因芯片；通过特异性抗体库与鼻咽癌肿瘤组织结合后，使标记引物扩增的V-D-J序列与抗体基因芯片进行杂交后显示癌组织的蛋白质表达谱，从而选择性通过免疫组织化学显示其中关键性蛋白质原位表达状况。结果 肿瘤组织结合的抗体滴度可达到106～108数量级，标记的DNA分子效价约为10-6～10-8，以该文一次检测20个抗体为例，基因芯片检出有10～14个阳性蛋白质表达谱；免疫组织化学可显示阳性抗体在细胞中的结合部位。结论 以抗体基因芯片为核心的技术方案，能在组织切片上同时检测含多个蛋白质表达谱,可以广泛应用于基础研究和临床诊疗过程。 |
| 关键词: 抗体基因芯片 蛋白质表达谱 噬菌体抗体库 |
| DOI：10.3969/j.issn.1674-3806.2012.11.02 |
| 分类号:R 73 |
| 基金项目:广西自然科学基金资助项目（编号：0728087） |
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| Antibody gene chip: a new method applied to detection of protein expression profile in biopsies of nasopharyngeal carcinoma |
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ZHU You-kai,SI Yong-feng,XIAO Rui-ping,et al.
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Department of Pathology,the People′s Hospital of Guangxi Zhuang Autonomous Region,Nanning 530021,China
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| Abstract: |
| ［Abstract］ Objective To develop an antibody gene chip to detect specific protein expression profile of nasopharyngeal carcinoma(NPC) tissues.Methods The specific antibody library containing anti-NPC antibodies were selected by using phage anti- body library. The V-D-J fragments as mark DNA molecules were used for preparation of antibody gene chip. After a number of different antibodies of specific antibody library binding the NPC tissues, the mark DNA molecules from the antibodies were amplified by PCR and the results of hybridization between the antibody gene chip and mark DNA molecules revealed protein expression profile of NPC tissues. The expression of key protein in situ of NP cells could be revealed by immunohisto- chemistry.Results The concentration degree of combinative antibodies could be up to 106～108. The mark DNA molecule titer could reach about 10-6～10-8；Gene chip detected 10～14 positive antibodies in 20 protein expression profile. The positions of binding antibodies in cells were showed by immunohistochemistry.Conclusion The antibody gene chip could detect protein expression profile in biopsies of NPC and could be widely used in basic research and clinical diagnosis and treatment process. |
| Key words: Antibody gene chip Protein expression profile Phage antibody library |
