摘要
目的制备SLC35F2多肽抗体，鉴定其性能，检测其对应蛋白在非小细胞肺癌（NSCLC）组织中的表达。方法通过蛋白质序列分析，选4段氨基酸序列合成多肽，混合免疫家兔，制备抗SLC35F2多肽抗体，经纯化、效价检测、特异性鉴定后，将抗体用于NSCLC标本免疫组织化学染色。结果获得兔抗入SLC35F2多肽抗体，效价1：10^5，Westernblot可特异识别41KD的SLC35F2蛋白，免疫组织化学示细胞胞质特异性染色。129例NSCLC的组织微阵列免疫组织化学染色显示，SLC35F2蛋白表达在90．7％（117／129）肺癌标本为廿到卅；而仅在17．1％（22／129）癌旁正常肺组织为廿，其余为-或＋；SLC35F2蛋白表达在NSCLC组织中高于癌旁正常肺组织（P〈0．01）；NSCLC病理分期与SLC35F2的升高表达存在低度相关（r=0．175）。结论用多肽免疫家兔制备抗SLC35F2抗体具有较高的效价和特异性，SLC35F2在NSCLC中表达高于癌旁正常组织。
Objective To prepare, purify and characterize the polyclonal antibody against SLC35F2, and detect the expression of SLC35F2 protein in non-small-cell lung cancer （NSCLC）. Methods Four polypeptides named peptide 1,2,3 and 4 were synthesized based on the bioinfformatics analysis of SLC35F2. The mixed polypeptides were injected into New Zealand rabbits. The polyclonal antibody was purified by immunoaffinity chromatography, and identified by Western blot and immunohistochemistry. The expression of SLC35F2 protein in 129 cases of NSCLC and corresponding adjacent normal lung tissues was detected by tissue microarray with immunohistochemistry. Results The titers of polyclonal antibody were 1：10^5 detected by ELISA assay. Western blot confirmed its high specific 41 KD band. The expression of SLC35F2 protein was strongly positive （ ＋＋ to ＋＋＋ ） in 90.7% （117/129） of the NSCLC samples. On the contrast ,only 17.1% （22/129） adjacent normal lung tissues were strongly positive （ ＋＋ ）. The expression of SLC35F2 protein was significantly higher in NSCLC than in the adjacent normal lung tissues （ P 〈 0.01 ）. Correlation analysis revealed that pathological stage had a low correlation with the expression of SLC35F2 protein. Factor analyses showed that 93.3% expression of SLC35F2 protein in NSCLC tissues were correlated with tumor differentiation and stage. Conclusion The specific antibody against human SLC35F2 was obtained. SLC35F2 protein was highly expressed in most of the NSCLC.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2009年第8期1046-1048,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（30672084）
关键词
非小细胞肺癌
人类溶质载体家族35成员F2
多克隆抗体
Non small celllung cancer
Homo sapiens solute carrier family 35 member F2
Polyelonal antibody
