期刊文献+

小鼠肿瘤坏死因子-α光激化学发光免疫法的建立 被引量:1

Establishment of Light Initiated Chemiluminescence Assay of Mouse TNF-α
下载PDF
导出
摘要 采用光激化学发光免疫法(LICA)建立快速定量检测小鼠mTNF-α。用两株mTNF-α不同表位的单克隆抗体(McAb),一株McAb包被发光微粒,另一株为生物素化McAb,两者与链亲和素包被的感光微粒构建双McAb夹心法mT-NF-α-LICA。数据采用双对数函数处理程序。结果表明:方法灵敏度为0.5ng/L;ED20、ED50、ED80分别为14.96 ng/L、77.5 ng/L、401.0 ng/L;可测范围为0.5~1200 ng/L;批内和批间CV分别为7.8%和10.5%;平均回收率为100.3%。mTNF-α-LICA与ELISA有较好的相关性。本文建立的mTNF-α-LICA法是mTNF-α检测中灵敏的方法,该方法稳定性好,具有很好的应用前景。 To establish a mouse TNF-α level, two quantitative light initiated chemiluminescence assay (LICA) for the determination of specific McAbs were selected. One was coated on iuminescence particles and the other was conjugated with biotin. These two McAb with sensitizer particles which were coated with streptavidin were used to constructed mTNF-α-LICA system. The results showed that the analytical sensitivity was 0.5ng/L. The 20% , 50% and 80% inhibition binding effect dose were 14.96 ng/L, 77.5 ng/L and 401.0ng/L,respectively. The detection range was from 0.5 ng/L to 1200 ng/L. The intra- and inter- assay CV of mTNF-α-LICA were 7.8% and 10.5% ,respectively, and the mean recovery rate was 100.3%. The results detected with mT- NF-et-LICA were well correlated with ELISA. The mTNF-α-LICA established in this study was a sensitive, sim- ple and convenient method in detecting mTNF-α level. The mTNF-α-LICA with high sensitivity and wide optimal range showed a great application prospect.
出处 《标记免疫分析与临床》 CAS 2009年第3期160-162,共3页 Labeled Immunoassays and Clinical Medicine
关键词 光激化学发光免疫法 肿瘤坏死因子-Α 免疫测定 单克隆抗体 小鼠 Light initiated chemiluminescence assay TNF-α Immunoassay McAb Mouse
  • 相关文献

参考文献8

  • 1Awasthi G,Singh S,Dash A P,et al.Genetic characterization and evolutionary inference of TNF-alpha through computational analysis[J].Braz J Infect Dis,2008,12(5):374-379.
  • 2Figiel I,Dzwonek K.TNF alpha and TNF receptor 1 expression in the mixed neuronal-glial cultures of hippocampal dentate gyrus exposed to glutamate or trimethyltin[J].Brain Res,2007,1131(1):17-28.
  • 3刘小伟,卢放根,游宇,练光辉,欧阳春晖,羊东晔,吴小平.TNF-α为中心的肝细胞凋亡与LPS所致小鼠肝损伤的关系[J].医学临床研究,2005,22(8):1051-1054. 被引量:3
  • 4Carswell E A,Old L J,Kassel R L,et al.An endotoxin-induced serum factor that causes necrosis of tumors[J].Proc Nat Acad Sci USA,1975,72(9):3666-3670.
  • 5Himmerich H,Fulda S,Linseisen J.TNF-alpha,soluble TNF receptor and interleukin-6 plasma levels in the general population[J].Eur Cytokine Netw,2006,17(3):196-201.
  • 6Haagmans B L,Stals F S,van der Meide P H,et al.Tumor necrosis factor alpha promotes replication and pathogenicity of rat cytomegalovirus[J].J Virol,1994,68(4):2297-2304.
  • 7Kouwenhoven M,Ozenci V,Teleshova N,et al.Enzyme-linked immunospot assays provide a sensitive tool for detection of cytokine secretion by monocytes[J].Clin Diagn Lab Immunol,2001,8(6):1248-1257.
  • 8李梅,宋达琳,康维强.细胞因子检测技术进展[J].国际检验医学杂志,2006,27(6):540-542. 被引量:8

二级参考文献22

  • 1Manuel PR, Josefa R, Manuel MR. et al . Vascular endothelial growth factor production in peritoneal macrophages of cirrhotic patients: regulation by eytokines and bacterial lipopolysaccharide[J].Hepatology . 1999,29:1057-1063.
  • 2Takehiko U, Matthias F, Gavin EA, et al . Toll like receptor 4 is involved in the mechanism of early alcohol-induced liver injury in mice[J]. Hepatology , 2001,34:101-108.
  • 3Vishnyakova TG, Bocharov AV. Baranova IN. et al . Binding and internalization of lipopolysaccharide by Cla-1, a human orthologue of rodent scavenger receptor B1[J]. J Biol Chem , 2003,278(25) :22771-22780.
  • 4Michael DJ, Rena FB, Kunitaro F, et al . Lipopolysaccharide and D-galactosamine-induced hepatic injury is mediated by TNF-a and not by Fas ligand[J]. Am J Physiol Regulatory Integrative Comp Physiol , 2000, 278 : R1196-R1201.
  • 5Schoemaker MH, Ros JE, Homan M, et al . Cytokine regulation of pro- and anti-apoptotic genes in rat hepatocytes: NF-kappaB-regulated inhibitor of apoptosis protein 2 (clAP2)prevents apoptosis[J]. J Hepatol , 2002,36(6):742-750.
  • 6Zhao Y, Li S, Childs EE, et al . Activation of pro-death Bcl-2 family proteins and mitochondria apoptosis pathway in tumor necrosis factor-alpha-induced liver injury[J]. J Biol Chem , 2001, 276(29):27432-27440.
  • 7Charles CB, Eva XB, Patricia EG, a al . Bacterial lipopolysaccharide enhances aflatoxin B1 hepatotoxicity in rat.s by a mecha-nism that depends on tumor necrosis factor α[J]. Hepatology ,2001 ;33:66-73.
  • 8Oberholzer A, Oberholzer C, Bahjat FR, et al. Genetic determinants of lipopolysaccharide and D-galactosamine-mediated hepatocellular apoptosis and lethality[J].J Endotoxin Res , 2001 ;7(5):375-380.
  • 9Murohisa G, Kohayashi Y, Kawasaki T, et al . Involvement of platelet-activating factor in hepatic apoptosis and necrosis inc hronic ethanol-fed rats given endotoxin[J]. Liver , 2002;22(5) :394-403.
  • 10Garcia-Ruiz C, Colell A, Mari M, et al. Defective TNF-alpha mediated hepatocellular apoptosis and liver damage in a cidic sphingomyelinase knockout mice [J].J Clin Invest 2003,111(2):197-208.

共引文献9

同被引文献6

  • 1盛建武,何苗,施汉昌,钱易.微囊藻毒素免疫检测技术研究进展[J].免疫学杂志,2005,21(B06):9-12. 被引量:4
  • 2盛建武,何苗,施汉昌,钱易.水环境中微囊藻毒素检测技术研究进展[J].环境污染与防治,2006,28(2):132-136. 被引量:22
  • 3Waller H, Chatterji U, Gallay P, et al. The use of AlphaLISA technology to detect interaction between hepatitis C virus-encoded NSSA and cyclophilin A [J]. J Virol Methods, 2010,165 : 202-210.
  • 4Cauchon E, Liu S, Percival MD, et al. Development of a homogeneous immunoassay for the detection of angiotensin I in plasma using AlphaLISA acceptor beads technology [J]. Anal Biochem,2009,388: 134-139.
  • 5McGiven JA, Sawyer J, Perrett LL, et al. A new homogeneous assay for high throughput serological diagnosis of brucellosis in ruminants [J]. J Immunol Methods, 2008,337 : 7-15.
  • 6蔡冲,李志凌.饮用水中微囊藻毒素检测研究进展[J].现代预防医学,2008,35(6):1140-1144. 被引量:3

引证文献1

二级引证文献2

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部