BLI技術(shù)應(yīng)用新型生物制品快速定量技術(shù)—分析

1、Better Lives. Better Planet.SMThis presentation is the Confidential work product of Pall Corporation and no portion of this presentation may be copied, published, performed, or redistributed without the express written authority of a Pall corporate officer 2016 Pall Corporation新型生物制品快速定量新型生物制品快速定量技技

2、術(shù)術(shù)BLI 技技術(shù)應(yīng)術(shù)應(yīng)用用China User Meeting2016-4-8陳陳濤濤2Agenda 直接定量法直接定量法 抓取定量法抓取定量法 雙雙抗夾心法（無(wú)信號(hào)放大）抗夾心法（無(wú)信號(hào)放大） 雙抗夾心法（有信號(hào)放大）雙抗夾心法（有信號(hào)放大） 競(jìng)競(jìng)爭(zhēng)爭(zhēng)法（無(wú)信號(hào)放大）法（無(wú)信號(hào)放大） 競(jìng)競(jìng)爭(zhēng)法（有信號(hào)放大）爭(zhēng)法（有信號(hào)放大）3使用BLI技術(shù)進(jìn)行濃度定量Relative IntensityWavelength (nm)100%0nm shiftTime傳感器起始斜率與濃度呈正比。分析物測(cè)試在幾分鐘內(nèi)完成。可以檢測(cè)粗樣品直接測(cè)定非標(biāo)記分析物的結(jié)合！4直接定量法 -analyteproA/pro

3、G/proL/AMQ/AHQ/GST/HISIgG,GST/his tagInitial slope檢測(cè)濃檢測(cè)濃度范度范圍圍:0.1ug/ml-200ug/ml 重復(fù)性重復(fù)性:10 times；（SA/AR2G）視固化穩(wěn)定性而定耗耗時(shí)時(shí):1-2 min/8 samples 軟軟件匹配件匹配:可以是否需要其他是否需要其他試劑試劑:不 計(jì)計(jì)算方法算方法:起始斜率應(yīng)應(yīng)用用領(lǐng)領(lǐng)域：域：蛋白，病毒的定量或者做Customized biosensor:SA/AR2G 固化抗體5直接定量法舉例-使用proA傳感器定量hIgG 6抓取定量法-antibody-analyteproA/proGProtein/v

4、irusInitial slope定量范定量范圍圍:0.1ug/ml-500ug/ml 重復(fù)性重復(fù)性:10 times 時(shí)間時(shí)間:10 min/8 samples 軟軟件兼容性件兼容性:yes需要的其他需要的其他試劑試劑:抗體或者受體 分析方法分析方法:initial slope應(yīng)應(yīng)用用:蛋白，疫苗類樣品的快速定量 Regeneration，pH1.7 glycine7抓取定量法舉例-某種病毒疫苗Lot 1Lot 2Step TypeAssay Time(s)solutionBaseline60Antibody bufferLoading30020ug/ml Ab(3ug/ml for 2Ga

5、)Baseline60antigen bufferAssociation180antigen dilutions8雙抗夾心法（無(wú)信號(hào)放大）定量范定量范圍圍:0.001ug/ml-10ug/ml 重復(fù)性重復(fù)性:10 times(not applied to SA biosensor) 時(shí)間時(shí)間:15-20 min/8 samples 軟軟件是否匹配件是否匹配:yes 分析方法分析方法:R equilibrium需要的其他需要的其他試劑試劑:one binding partner(virus/saccharide),two binding partner(monomer protein)應(yīng)應(yīng)用用:

6、生物大分子的快速定量 proA biosensor-Ab-analyte-HIgG block-AbRegeneration，pH1.7 glycineSA biosensor-Ab-analyte-Abvirus/saccharideR equilibrium9雙抗夾心法（無(wú)信號(hào)放大）舉例-某種疫苗定量Ab loading vaccine bindingHIgG blockAb binding20ug/ml2ug/ml0.2ug/ml0.02ug/ml0.002ug/ml20ug/ml2ug/ml0.2ug/ml0.02ug/ml0.002ug/ml10雙抗夾心法（有信號(hào)放大）定量范圍:0.

7、001ug/ml-1ug/ml 重復(fù)性重復(fù)性:5-10%再生次數(shù)再生次數(shù):no regenerable實(shí)驗(yàn)時(shí)間實(shí)驗(yàn)時(shí)間:30 min/8 samples 軟軟件是否匹配件是否匹配:yes 分析方法分析方法:R equilibrium需要的其他需要的其他試劑試劑:one binding partner(virus/saccharide),two binding partner(monomer protein),Enzyme,DAB 應(yīng)應(yīng)用用:residual contamination,PK,diagnosis kitSA biosensor-Ab-analyte-2nd Ab-enzyme-D

8、ABProtein/virus/saccharideR equilibrium11雙抗夾心法（有信號(hào)放大）舉例-CHO HCP kit12競(jìng)爭(zhēng)法（無(wú)信號(hào)放大）APS/AR2G-analyte-analyte mixed with AbSmall moleculeHigh concentrationlow concentration檢測(cè)檢測(cè)方位方位:1ng/ml-1000g/ml 重復(fù)性重復(fù)性:5-10%再生能力再生能力:no regenerable時(shí)間時(shí)間:30 min/8 samples軟軟件是否匹配件是否匹配:分析軟件沒(méi)有競(jìng)爭(zhēng)模式需要的其他需要的其他試劑試劑:one binding par

9、tner,Enzyme,DAB 應(yīng)應(yīng)用用:高靈敏度小分子濃度測(cè)試13競(jìng)爭(zhēng)法（無(wú)信號(hào)放大）舉例-抗生素殘留測(cè)試Unlabled-Unlabled-BLIBLI detect detect Ampicillin(Without Nano Gold Ampicillin(Without Nano Gold Particles)Particles)Labled-Labled-BLIBLI detect Ampicillin(With detect Ampicillin(With Nano Gold Particles)Nano Gold Particles)R Conc.Dynamic rangeYes/No cut off value14競(jìng)爭(zhēng)法（有信號(hào)放大）檢測(cè)檢測(cè)方位方位:0.1ng/ml-1000g/ml 重復(fù)性重復(fù)性:5-10%再生能力再生能力:no regenerable時(shí)間時(shí)間:30 min/8 samples軟軟件是否匹配件是否匹配:分析軟件沒(méi)有競(jìng)爭(zhēng)模式需要的其他需要的其他試劑試劑:one binding partner,Enzyme,DAB 應(yīng)應(yīng)用用:高靈敏度小分子濃度測(cè)試APS biosensor-analyte-a