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	脑钠肽单克隆抗体的制备及临床应用研究; Preparation and selection of the monoclonal antibody used for kits to detect BNP32 which was applied to clinical research
	何涛君 ; 陈洲 ; 邱竹英 ; 王琼 ; 陆学东
	2010
关键词	脑钠肽 单克隆抗体 心衰 ELISA brain natriuretic peptide(BNP) monoclonal antibody heart failure ELISA
英文摘要	目的:制备抗脑钠肽(bnP32)单克隆抗体(MAb),并利用双抗体夹心ElISA法建立bnP抗原检测技术,应用于临床心脏患者脑钠肽水平的检测。方法:以基因工程原核重组表达bnP抗原免疫bAlb/C小鼠,利用常规杂交瘤技术制备MAb,MAb经纯化和HrP标记后,利用双抗体夹心ElISA法筛选检测bnP32蛋白的最佳配对MAb,以其建立bnP32抗原检测技术,并与临床bnP检测的标准实验做平行比较。结果:成功筛选到16株稳定分泌抗bnP32MAb的杂交瘤细胞株,16株MAb的亚型分别为Igg1、Igg2A和IgM,并从中筛选出最佳MAb配对组合,该组合对bnP32蛋白的检测灵敏度为20ng/l。建立的双抗体夹心bnP检测ElISA法与临床bnP检测的标准实验平行比较具有很好的一致性(kAPPA值=0.828),两者没有统计学意义(P>0.05)。结论:成功地建立了bnP32抗原的双抗体夹心ElISA法检测技术,并能够很好地运用于临床心衰患者bnP指标的检测。; AIM:To prepare the monoclonal antibody(mAb)used for kits to detect the BNP32 antigen by means of double-antibody sandwich ELISA assay.Comparasion differences on detection BNP between double-antibody sandwich ELISA and Roche ECL.METHODS:BALB/c mice were immunized with purified recombinant BNP32 protein and by routine hybridoma technique,Then comparasion differences on detection BNP between double-antibody sandwich ELISA and Roche ECL.RESULTS:16 hybridoma cell lines secreting potent mAb against BNP32 were obtained.The subtype of these 16 mAb were found to be IgG1,IgG2a and IgM,then their cross-blocking properties were analyzed,when they reacted with the BNP32 protein in direct ELISA in order to offer the valuable data for selecting feasible pair of mAb in the detection of the BNP32 antigen.All the mAbs were used for the detection of BNP32 by double-antibody sandwich ELISA.In addition,the mAbs were purified and HRP-labelled in advance.Finally,we had successfully screened a pair of mAbs,which exhibited a sensitivity of 20 ng/L for the detection of BNP32 antigen.The two detection methods have very good consistency(kappa=0.828>0.75)between double-antibody sandwich ELISA and Roche ECL.There was no statistics significance on differences between these two methods(P>0.05).CONCLUSION:It is evident that this pair of mAb shows excellent detection of BNP32.The double-antibody sandwich ELISA can be good apply to detect BNP level in clinical congestive heart failure patients.
语种	zh_CN
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/119006]
专题	生命科学－已发表论文
推荐引用方式 GB/T 7714	何涛君,陈洲,邱竹英,等. 脑钠肽单克隆抗体的制备及临床应用研究, Preparation and selection of the monoclonal antibody used for kits to detect BNP32 which was applied to clinical research[J],2010.
APA	何涛君,陈洲,邱竹英,王琼,&陆学东.(2010).脑钠肽单克隆抗体的制备及临床应用研究..
MLA	何涛君,et al."脑钠肽单克隆抗体的制备及临床应用研究".(2010).

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