目的建立轉(zhuǎn)基因細(xì)胞法測(cè)定抗CD52單克隆抗體(單抗)的抗體依賴(lài)細(xì)胞介導(dǎo)的細(xì)胞毒性作用(antibody-dependent cell-mediated cytotoxicity, ADCC)生物學(xué)活性檢測(cè)方法。方法①以Ramos細(xì)胞系作為靶細(xì)胞,以Jurkat-hFcγRⅢa/FcεRIγ-NFAT轉(zhuǎn)基因細(xì)胞系作為效應(yīng)細(xì)胞,通過(guò)熒光素酶檢測(cè)系統(tǒng)(Bright-Glo^TM luciferase Assay System)建立抗CD52單抗的ADCC生物學(xué)活性檢測(cè)方法;②對(duì)靶細(xì)胞、量效范圍、效靶比及誘導(dǎo)時(shí)間進(jìn)行優(yōu)化并進(jìn)行方法學(xué)驗(yàn)證;③研究建立的方法應(yīng)用于對(duì)不同抗CD52單抗的ADCC生物學(xué)活性檢測(cè)。結(jié)果建立抗CD52單抗的ADCC生物學(xué)活性檢測(cè)方法,抗CD52單抗在該方法中存在量效關(guān)系,符合四參數(shù)方程:y=(A-D)/[1+(x/C)^B]+D;經(jīng)優(yōu)化后確定抗體量效范圍為起始質(zhì)量濃度360μg/mL,4倍系列稀釋9個(gè)稀釋度;效靶比為3∶1,誘導(dǎo)時(shí)間為6.0 h;該方法具有良好的專(zhuān)屬性;4個(gè)不同稀釋組回收率樣品經(jīng)3次測(cè)定,相對(duì)效價(jià)分別為(45.58±...

【文章頁(yè)數(shù)】：7 頁(yè)

【部分圖文】：

圖1抗CD52單抗的ADCC劑量效應(yīng)曲線Fig．1Dose-responsecurveofADCCpotencyofanti-CD52mAb

圖1抗CD52單抗的ADCC劑量效應(yīng)曲線Fig．1Dose-responsecurveofADCCpotencyofanti-CD52mAb圖2ADCC中不同靶細(xì)胞對(duì)比Fig．2ComparisonofdifferenttargetcellsinADCCassay2．2．2抗體量....

圖2ADCC中不同靶細(xì)胞對(duì)比Fig．2ComparisonofdifferenttargetcellsinADCCassay

圖1抗CD52單抗的ADCC劑量效應(yīng)曲線Fig．1Dose-responsecurveofADCCpotencyofanti-CD52mAb圖2ADCC中不同靶細(xì)胞對(duì)比Fig．2ComparisonofdifferenttargetcellsinADCCassay2．2．2抗體量....

圖3ADCC中單抗量效范圍的優(yōu)化Fig．3Optimizationofdose-responsecurveofanti-CD52

圖1抗CD52單抗的ADCC劑量效應(yīng)曲線Fig．1Dose-responsecurveofADCCpotencyofanti-CD52mAb圖2ADCC中不同靶細(xì)胞對(duì)比Fig．2ComparisonofdifferenttargetcellsinADCCassay2．2．2抗體量....

圖4ADCC中不同效靶比對(duì)比Fig．4Comparisonofdifferentratioofeffectortotarget

圖1抗CD52單抗的ADCC劑量效應(yīng)曲線Fig．1Dose-responsecurveofADCCpotencyofanti-CD52mAb圖2ADCC中不同靶細(xì)胞對(duì)比Fig．2ComparisonofdifferenttargetcellsinADCCassay2．2．2抗體量....

本文編號(hào)：3957748