光激化学发光技术介绍PPT课件.ppt

Click to edit Master title style.,Click to edit Master text styles,Second level,Third level,Fourth level,Fifth level,*,Biopharma 01.09.03,L,ight,I,nitiated,C,hemiluminescence,A,ssay,光激化学发光（,LiCA,）,1,Biopharma 01.09.03,光激化学发光（,LiCA,）介绍,以高分子微粒为基础,两种微粒近距离,离子氧能量传递,主要特点：,高灵敏度,低本底,稳定,均相,免洗,广泛应用于生物分子间的相互作用,2,Biopharma 01.09.03,微粒的组成,感光微粒内含感光化合物,发光微粒内含发光化合物和镧系元素,微粒直径,188nm,，表面覆盖水凝胶,共价连接,增大的表面积,成百上千个生物分子，捕获目标分子,3,Biopharma 01.09.03,LiCA,发光原理,核心原理是能量的近距离转移,转移的介质是高能态离子氧,红色激光,680nm,照射，感光微粒释放离子氧（,4,微秒），传播直径,200nm,，发光微粒（,520,618nm,），高能级的光,反应的基础有赖于两种微粒的相互接近,反应体系中，微粒的浓度很低，碰撞机率很低，本底信号微弱,4,Biopharma 01.09.03,原理,(1),发射光,520-620 nm,发光微球,1,O,2,A,B,激发光,680 nm,感光微球,200nm,5,Biopharma 01.09.03,原理,(2),激发光,680 nm,感光微球,A,发光微球,C,O,2-,6,Biopharma 01.09.03,LiCA,的技术特点（一）,高灵敏度,信号产生逐级放大的结果,每个感光微粒释放出近,6,万个离子氧,离子氧作用于二甲基噻吩，产生大量紫外光,紫外光激发镧系元素,实际检测能力为,10,-15,摩尔,理论检测能力为,10,-18,摩尔,7,Biopharma 01.09.03,LiCA,的技术特点（二）,低本底,时间分辨模式采集光信号（延迟,150 ms,），天然荧光的寿命小于,100 ns,采用,680 nm,红光激发，生物样品中的荧光物质不敏感,发射光的波长比激发光的波长短，能量更高，有别于荧光，有效避免荧光物质干扰,8,Biopharma 01.09.03,稳定,均相环境使反应更充分,免洗操作杜绝随机误差和交叉污染,感光和发光的有机分子受到微粒的保护，环境稳定和安全,溶液中的氧浓度是一个常数,不易受到,pH,值、离子强度和温度的影响,LiCA,的技术特点（三）,9,Biopharma 01.09.03,临床免疫的各项检测,生命科学的基础研究,新药的筛选,标记简便,检测过程简单,广泛应用,10,Biopharma 01.09.03,基本包被模式,GG-Streptavidin,（,SA,）,FG-Antibody,（,Ab,）,11,Biopharma 01.09.03,基本反应模式,biotin-antigen,primary Ab,secondary Ab(or protein A,G,LA),12,Biopharma 01.09.03,应用类型,配体受体,激酶,蛋白蛋白,免疫,蛋白酶,蛋白,DNA,13,Biopharma 01.09.03,配体受体,(1),TNFa-sTNFR1,Biotin,anti-TNFR1 IgG,FG,-TNFa,Protein A-,sTNFR1,14,Biopharma 01.09.03,配体受体,(1),TNFa-sTNFR1,GG-SA,Biotin,anti-TNFR1 IgG,FG,-TNFa,Protein A-,sTNFR1,15,Biopharma 01.09.03,配体受体,(1),TNFa-sTNFR1,GG-SA,Biotin,anti-TNFR1 IgG,FG,-TNFa,Protein A-,sTNFR1,16,Biopharma 01.09.03,配体受体,(2),环磷酸腺苷,cAMP,Biotin,FG,-cAMP,anti-cAMP-,17,Biopharma 01.09.03,配体受体,(2),环磷酸腺苷,cAMP,GG-SA,Biotin,FG,-cAMP,anti-cAMP-,18,Biopharma 01.09.03,激酶,Akt,Biotin,anti-Gsk3 IgG,FG,-Gsk3,Protein A-,19,Biopharma 01.09.03,激酶,Akt,Biotin,anti-Gsk3 IgG,FG,-Gsk3,Protein A-,20,Biopharma 01.09.03,激酶,Akt,GG-SA,Biotin,anti-Gsk3 IgG,FG,-Gsk3,Protein A-,Akt,磷酸化,21,Biopharma 01.09.03,激酶,Akt,GG-SA,Biotin,anti-Gsk3 IgG,FG,-Gsk3,Protein A-,Akt,22,Biopharma 01.09.03,蛋白蛋白,Biotin,FG,-p53,Anti-GST-,p53/HDM2,HDM2-,GST,23,Biopharma 01.09.03,蛋白蛋白,GG-SA,Biotin,FG,-p53,Anti-GST-,p53/HDM2,HDM2-,GST,24,Biopharma 01.09.03,免疫,(1),肿瘤坏死因子,TNFa,Biotin,FG,-anti-TNFa,anti-TNFa-,TNFa,25,Biopharma 01.09.03,免疫,(1),肿瘤坏死因子,TNFa,GG-SA,Biotin,FG,-anti-TNFa,anti-TNFa-,TNFa,26,Biopharma 01.09.03,免疫,(2),Biotin,anti-TNFa,FG,-anti-TNFa,Protein A-,TNFa,肿瘤坏死因子,TNFa,GG-SA,27,Biopharma 01.09.03,免疫,(2),GG-SA,Biotin,anti-TNFa,FG,-anti-TNFa,Protein A-,TNFa,肿瘤坏死因子,TNFa,28,Biopharma 01.09.03,蛋白酶,(1),GG-SA,Biotin,FG,-substrate,anti-digoxigenin-,组织蛋白酶,Cathepsin D,-digoxigenin,Cathepsin D,水解,29,Biopharma 01.09.03,蛋白酶,(2),ACE,Biotin,FG,-angiotensin I,anti-angiotensin II-,ACE,切割,30,Biopharma 01.09.03,蛋白酶,(2),ACE,GG-SA,Biotin,FG,-,angiotensin I I,anti-angiotensin II-,31,Biopharma 01.09.03,蛋白,DNA,ERa-DNA,Biotin,FG,-ERa,anti-digoxigenin-,-digoxigenin,DNA,32,Biopharma 01.09.03,蛋白,DNA,ERa-ERE,GG-SA,Biotin,FG,-ERa,anti-digoxigenin-,-digoxigenin,DNA,33,Biopharma 01.09.03,当前成功应用实例,34,Biopharma 01.09.03,HBsAg,GG-SA,Biotin,-Anti-HBs,Anti-HBs-FG,HBsAg,35,Biopharma 01.09.03,Anti-HBs,GG-SA,Biotin-HBsAg,FG,HBsAg-,Anti-HBs,36,Biopharma 01.09.03,HBeAg,GG-SA,Biotin,FG,-Anti-HBe,Anti-HBe-,HBeAg,37,Biopharma 01.09.03,Anti-HBe,GG-SA,Biotin,FG,-Anti-HBe,Anti-HBe-,中和,HBeAg,Sample Anti-HBe,38,Biopharma 01.09.03,Anti-HBc,GG-SA,Biotin,FG,-HBcAg,Anti-HBc-,Anti-HBc,39,Biopharma 01.09.03,检测平台,40,Biopharma 01.09.03,LiCA SP,双加样臂,气置换式微量进样泵,液面自动检测功能,标本、试剂装载,100,个标本,10,个试剂位,41,Biopharma 01.09.03,LiCA HT,一次装载,300,个测试,自动高速进样：,0.5,秒,/,孔,自动温育、震荡和读数,300,个测试仅需,50,分钟,42,Biopharma 01.09.03,谢 谢！,43,Biopharma 01.09.03,