| 地高辛标记核酸探针检测重组人白细胞介素1受体颉颃剂中DNA残余量的研究 |
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| 引用本文： | 王超,刘庚,张林波. 地高辛标记核酸探针检测重组人白细胞介素1受体颉颃剂中DNA残余量的研究[J]. 中国畜牧兽医, 2013, 40(11): 54-58 |
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| 作者姓名： | 王超 刘庚 张林波 |
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| 作者单位： | 1. 吉林农业大学生命科学学院, 吉林长春 130118;2. 吉林农业大学中药材学院, 吉林长春 130118 |
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| 基金项目： | 国家自然基金(31072127)。 |
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| 摘 要： | 试验旨在建立重组人白细胞介素1受体颉颃剂(interleukin-1 receptor antagonist,IL-1ra)原液中宿主菌DNA残余量的检测方法。以重组白细胞介素1受体颉颃剂工程菌基因组DNA为模板,优化试验条件,借助超声波手段,制备地高辛标记的核酸探针, 并以此探针进行狭缝杂交及显色反应。结果显示,在24 批次重组人IL-1ra原液中,每人份使用剂量的宿主DNA残余量均小于10 ng。结果表明,该方法灵敏度较高、特异性较强、重复性较好、操作步骤安全且较为方便,可用于重组人IL-1ra原液及半成品的DNA残余检测。
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| 关 键 词： | DNA 残余量 地高辛 重组人白细胞介素1受体颉颃剂 狭缝杂交 |
| 收稿时间： | 2013-03-28 |
Detection of Residual DNA in Recombinant Interleukin-1 Receptor Antagonist(IL-1ra) Using Digoxigenin Labeled Probe |
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| WANG Chao,LIU Geng,ZHANG Lin-bo. Detection of Residual DNA in Recombinant Interleukin-1 Receptor Antagonist(IL-1ra) Using Digoxigenin Labeled Probe[J]. China Animal Husbandry & Veterinary Medicine, 2013, 40(11): 54-58 |
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| Authors: | WANG Chao LIU Geng ZHANG Lin-bo |
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| Affiliation: | 1. College of Life Sicence, Jilin Agricultural University, Changchun 130118, China;2. College of Chinese Medicine Material, Jilin Agricultural University, Changchun 130118, China |
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| Abstract: | The assay was aimed to establish a method for detecting the residual DNA in recombinant interleukin-1 receptor antagonist (IL-1ra) using digoxigenin labeled probe. By optimizing experimental conditions, with ultrasonic means,genomic DNA of engineered bacteria containing IL-1ra was labeled by digoxigenin,as a probe for slot blotting assay to dectect residual DNA in bulk products. The residual DNA in 24 batches of bulk products was less than 10 ng per dosage for per person. This method was sensitive, specific and convenient,and could be well repeated, which could be used for detecting residual DNA of recombinant IL-1ra protein bulk products and semi-finished products. |
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| Keywords: | residual DNA digoxigenin recombinant interleukin-1 receptor antagonist slot blotting |
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