摘要
目的生物信息学分析NAP-2(73),以及制备其多克隆抗体。方法生物信息学分析预测出NAP-2(73)蛋白质的等电点、亲水性、抗原性等理化特性,以这些分析预测为依据,设计出一段抗原多肽,免疫家兔得到多克隆抗体,并用ELISA、West-ernblot等方法鉴定其滴度和特异性。结果对NAP-2(73)的功能性位点、抗原性和亲水性等进行了预测分析,成功制备了多克隆抗体,其血清中抗NAP-2(73)抗体效价为1:729000,并以此抗体证实了NAP-2(73)在肝癌细胞和组织中的表达。结论利用生物信息学,高效价和特异性强的抗NAP-2(73)抗体被成功制备,为进一步研究奠定了基础。
NAP-2（73） is a proteomic biomarker of HCC.This study is aimed to analyze NAP-2（73） by means of bioinformatics and prepare its polyclonal antibody.We analyzed the peptide sequence of NAP-2（73） according to its bioinformatics,and synthesized the sequences on the basis of hydrophilicity,antigenicity,accessibility and flexibility.The synthesized peptide was used to immunize rabbits for production of Ab.And the titer of the prepared antibodies was analyzed by ELISA.We applied Western blotting to analyze NAP-2（73） expression in the HCC cells lines and in hepatocellular carcinoma tissues,while we also detected the adjacent tissues by immunohistochemical staining.In this study,the functional sites,hydrophilicity,antigenicity of NAP-2（73） were predicted by means of bioinformatics;the polyclonal antibody of NAP-2（73） was successfully obtained,and the titer of the antibody in serum was 1：729 000.The expressions of NAP-2（73） in the HCC cells lines and hepatocellular carcinoma tissues were conformed.Our results indicate that specific and high titer polyclonal antibody against NAP-2（73） can be prepared successfully by means of bioinformatics and provide a basis for further studying.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2010年第7期628-631,共4页
Immunological Journal
基金
广西自然科学基金资助项目(桂科攻0592007-1C
桂科能0630006-5E
桂科攻0993003C-13)
