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

Abstract

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.

Key words: residual DNA; digoxigenin; recombinant interleukin-1 receptor antagonist; slot blotting

CLC Number:

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.

References

1 刁有祥, 丁家波, 姜世金, 等. 地高辛标记探针检测猪胸膜肺炎放线杆菌的研究与应用[J]. 畜牧兽医学报, 2005,36(6): 585~589.
2 王军志.生物技术药物研究开发和质量控制[M].第2版.北京: 科学出版社,2007.
3 刘方娜, 刁有祥, 王妮, 等. 猪圆环病毒2型地高辛标记探针检测方法的建立与应用[J]. 中国兽医学报, 2008,28(7): 771~774.
4 刘志杰, 任慧英, 温建新,等.用地高辛标记的核酸探针检测猪伪狂犬病毒野毒感染的研究[J]. 畜牧兽医学报, 2008,39(11): 1621~1624.
5 佘清.第58届WHO生物制品标准化专家委员会会议简况[J].中国药品标准,2007,8(1):77.
6 李广明, 姜平, 李玉峰, 等. 地高辛标记cDNA探针检测猪繁殖与呼吸综合征病毒[J]. 畜牧与兽医, 2007,39(5): 43~45.
7 李加, 唐建蓉, 刘景华,等. 斑点杂交法测定Vero细胞DNA残余量的探针选择[J].药物分析杂志,2008,28(11):1892~1895.
8 李瑶瑶, 刁有祥, 李宏梅. 地高辛标记探针检测鼻气管鸟杆菌的研究与应用[J]. 西北农林科技大学学报(自然科学版), 2010,38(4): 19~23.
9 邵艳红, 李咏兰, 曹贵方,等. 原位杂交技术检测蒙古绵羊胎儿β-防御素的表达[J].中国畜牧兽医, 2008,35(9): 48~52.
10 国家药典委员会.中华人民共和国药典(三部)[M].第9版.北京:化学工业出版社,2010.
11 孟凡磊, 刁有祥, 王辉, 等. 鸡传染性支气管炎病毒地高辛标记探针检测方法的的建立和应用[J]. 福建农业学报, 2007,22(1): 39~42.
12 高绪慧,刁有祥, 唐熠,等. 探针检测鸭黄病毒的地高辛标记DNA的制备与应用[J]. 中国兽医学报, 2012,32(4): 525~528.
13 崔治中, 赵鹏, 孙淑红,等. 鸡致病性外源性禽白血病病毒特异性核酸探针交叉斑点杂交检测试剂盒的研制[J]. 中国兽药杂志, 2011,45(8): 5~11.
14 黄相红,胡立德,梁文璐,等. 地高辛标记探针检测重组人p43蛋白宿主DNA残余量的研究[J]. 药物分析杂志,2011,31(2):356~359.
15 韩先杰, 王君伟, 布日额,等. 地高辛标记核酸探针检测鸭瘟病毒[J]. 中国兽医杂志, 2004,40(11): 20~21.
16 Arend W P,Guthridge C J.Biological role of interleukin-1 receptor antagonist isoforms[J].Ann Rheum Dis,2000,59(Suppl):1160~1164.
17 Garnick R L, Solli N J, Papa P A. The role of quality control in biotechnology: An analytical perspective[J]. Anal Chem, 1988,60(23): 2546~2557.
18 Ji X, Lee K, DiPaolo B. High-sensitivity hybridization assay for quantitation of residual E.coli DNA[J]. Biotechniques, 2002, 32(5):1162.
19 Martins S A M, Prazeres D M F, Cabral M S, et al. Comparison of real-time polymerase chain reaction and hybridization assays for the detection of E.coli genomic DNA in process sample and pharmaceutical grade plasmid DNA products[J].Anl Biochem,2003,322(1):127.
20 Wang J Y, Yu Y, Feng L X, et al. Genomic DNA extraction from hair sacs of pigs using modified phenol-chloroform method[J]. Herediatas, 2010,32(7): 752~756.

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Detection of Residual DNA in Recombinant Interleukin-1 Receptor Antagonist(IL-1ra) Using Digoxigenin Labeled Probe

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