北华大学学报（自然科学版）2017，Vol.18Issue(5)：598-602,5.DOI:10.11713/j.issn.1009-4822.2017.05.008
P-5m八肽多克隆抗体的制备及应用
Preparation and Application of P-5m Octapeptide Polyclonal Antibodies
摘要
Abstract
Objective To prepare polyclonal antibody against P-5m octapeptide for preliminary identification and application, and to obtain important experimental tools for studying the function and mechanism of P-5m octapeptide. Method P-5m octapeptide was synthesized by 9-fluoromethoxycarbonyl ( Fmoc-) solid phase methods, purified and linked with carrier protein-keyhole limpet hemocyanin. The conjugated peptide was subcutaneously immunized with New Zealand white rabbits and booster immunized with the conjugated peptide, the anti-serum was isolated and purified by C18 reverse phase chromatography. The serum titer was determined by indirect ELISA. Transwell and western blot assays were used to detect the effect of polyclonal antibody antagonizing P-5m octapeptide on tumor metastasis. Results The titer of the purified antibody was more than 1 : 160000 by indirect ELISA, indicated that the high titer polyclonal antibody was obtained. Transwell experiments showed that the antibody can inhibit the inhibition of P-5m on tumor cell invasion. Western blotting method showed the polyclonal antibody was highly specific to P-5m octapeptide,and it could block the expression of E-cadherin and decrease the expression of MMP-2 and MMP-9 in SW620 cells, which was statistically significant ( P<0 . 05 ) . Conclusion The prepared P-5m polyclonal antibody could be used in transwell and western blot assays,and provided the experimental basis to find the function of P-5m protein and its mechanism of anti-tumor metastasis.关键词
P-5m八肽/多克隆抗体/结肠癌Key words
P-5m octapeptide/polyclonal antibody/colon carcinoma分类
生物科学引用本文复制引用
陶彤彤,韩笑,杨昕祺,侯志东,安丽萍,徐广宇,杜培革..P-5m八肽多克隆抗体的制备及应用[J].北华大学学报（自然科学版）,2017,18(5):598-602,5.基金项目
国家自然科学基金项目(31201061) （31201061）
吉林省教育厅科学技术研究项目(2015144). （2015144）
