本文關(guān)鍵詞： 雜交瘤細胞株 單克隆抗體 流腦疫苗　出處：《南京醫(yī)科大學學報(自然科學版)》2016年11期 　論文類型：期刊論文

【摘要】：目的 :篩選能分泌A群腦膜炎奈瑟球菌多糖抗體的雜交瘤細胞株,制備單克隆抗體,運用該單抗建立檢測ACYW135群腦膜炎奈瑟球菌多糖疫苗中A群多糖的競爭ELISA方法。方法 :運用經(jīng)典的雜交瘤技術(shù)篩選雜交瘤細胞株,采用小鼠腹腔注射收獲腹水型單克隆抗體,優(yōu)化條件建立間接競爭ELISA,分別測定5支同一批和3個不同批次的ACYW135群腦膜炎奈瑟球菌多糖疫苗成品中的A群多糖含量,檢測結(jié)果應(yīng)用SPSS軟件做單樣本t檢驗分析。結(jié)果:標準曲線經(jīng)Log-logit處理,得到回歸方程:y=-1.664 8-2.254 2x,R2=0.989 5,線性范圍0.022 8~1.200 7μg/m L,IC50為0.165 8μg/m L,最低檢測限為0.022 8μg/m L,檢測疫苗中A群多糖含量,檢測值與疫苗標示量無統(tǒng)計學差異(P0.05)。結(jié)論 :成功篩選雜交瘤細胞株,制備A群多糖特異性單克隆抗體,建立的間接競爭ELISA方法 ,靈敏度高、穩(wěn)定性好、可重復性強,可用于腦膜炎奈瑟球菌多糖疫苗研發(fā)過程中的定量檢測,能克服磷含量法和火箭免疫電泳法檢測A群多糖含量的缺點。
[Abstract]:Objective: to screen hybridoma cell lines that secrete antibodies against Neisseria meningitidis polysaccharide in group A and to prepare monoclonal antibodies. The monoclonal antibody was used to establish a competitive ELISA method for the detection of group A polysaccharides from Neisseria meningitidis polysaccharide vaccine in ACYW135 group. Methods: hybridoma cell lines were screened by classical hybridoma technique, and ascites monoclonal antibodies were harvested by intraperitoneal injection of mice. The optimized conditions were established to establish indirect competitive ELISA.The concentration of group A polysaccharides in the finished products of five ACYW135 group meningitis Neisseria meningitidis polysaccharides vaccine was determined respectively. Results: the standard curve was processed by Log-logit, and the regression equation was obtained: the regression equation was: Yi-1.664 8-2.2542xR2N 0.989 5. The linear range was 0.022 8 渭 g / m -1. 200 7 渭 g / L IC50 was 0.165 8 渭 g / mL, the lowest detection limit was 0.022 8 渭 g / mL, and the detection limit was 0.022 8 渭 g / mL. Conclusion: the hybridoma cell line was screened successfully, the specific monoclonal antibody against group A polysaccharides was prepared, and the indirect competitive ELISA method was established with high sensitivity, good stability and strong reproducibility. It can be used for quantitative detection of Neisseria meningitidis polysaccharide vaccine, and can overcome the shortcomings of phosphorus content and rocket immunoelectrophoresis.
【作者單位】： 昆明醫(yī)科大學暨云南省天然藥物藥理重點實驗室;沃森生物技術(shù)股份有限公司;
【基金】：云南省高新技術(shù)產(chǎn)業(yè)發(fā)展項目([2014]1264)
【分類號】：R392

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