强力霉素单克隆抗体制备与检测_生物工程.doc

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  • 更新时间:2014-11-03
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摘要:强力霉素作为一种抗生素,在日长生活中的大量不合理的使用滥用,导致兽药在食品中的大量残留,严重影响到人们的日常生活。如何快速检测这类抗生素已成为当务之急。目前对强力霉素的残留分析多采用微生物法、色谱法及色谱 -质谱联用法等。但这些方法耗时,而且测试费用高,前处理步骤繁杂, 不能满足速检测的要求。酶联免疫吸附测定法( ELISA )具有快速、 准确、 高通量的特点, 可以极大地提高检测效率,节约检测成本, 现已被广泛应用于食品安全快速检测中。

   本实验采用强力霉素半抗原经过人工合成抗原免疫小白鼠,通过ELISA检测小鼠血清效价为1:6400,选择效价值最高的小白鼠脾细胞和小鼠骨髓瘤细胞sp2/0并以选择培养基培养融合细胞,再通过ELISA检测筛选出阳性的杂交瘤细胞。适当克隆扩增得到具有特异分泌的单克隆杂交瘤细胞株。为强力霉素的及时,快速检测奠定了基础。

关键词 强力霉素;细胞融合;单克隆抗体;酶联免疫吸附实验

 

Abstract:Doxycycline as an antibiotic,In ordinary life, a large number of unreasonable use abuse,Lead to veterinary drugs in a large number of residues in food, seriously affecting people's daily lives.How to quickly detect such antibiotics, has become imperative.Currently, doxycycline residue analysis Use more, microbiological method, chromatography and chromatography - mass spectrometry, etc.However, these methods are time consuming Moreover, the high cost of testing, the complex pre-processing step,Can not meet the requirements of speed detection.Enzyme-linked immunosorbent assay (ELISA) is fast, accurate, high-throughput characteristics,Can greatly improve the detection efficiency, saving the cost of detection, has been widely used in the rapid detection of food safety.

   The present study, doxycycline hapten the synthetic antigen immune mice,Detected by ELISA The serum detection titer of 1:6400,Select the highest value of efficiency, mouse spleen cells and mouse myeloma cells SP2 / 0,Selective medium culture fusion cells,By ELISA detection screened positive hybridoma。Appropriately amplified cloned and obtained monoclonal hybridoma strain With a specific secretion

Keywords  Doxycycline ; Cell fusion; Monoclonal antibody; Synthesis Enzyme-linked immunosorbent assay


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