人白血病抑制因子受体单抗靶向脂质体微泡的制备及其体外荧光鉴定

目的 制备人白血病抑制因子受体( LIFR)单抗靶向脂质体微泡,并对其微泡微囊的连接可靠性进行体外免疫荧光鉴定.方法 利用生物素-亲和素桥连技术,构建LIFR单抗脂质体微泡(MB-BSB-LIFR-AB).以FITC荧光亲和素标记普通脂质体微泡(MB)、生物素化脂质体微泡(MB-B),以两个不同浓度梯度(1:4和1:16)的DTAF荧光二抗分别标记MB、MB-B、生物素-亲和素脂质体微泡(MB-BS)、MB-BSB-LIFR-AB;荧光显微镜下观察微泡荧光强度并分为0级、1级、2级、3级.结果 加入FITC荧光亲和素后,MB-B呈现3级明亮的绿色荧光,而MB无荧光显示(0级)；以1:4和1:16浓度梯度的DTAF荧光二抗孵育后,MB-BSB-LIFR-AB均发出3级明亮的绿色荧光；而MB-BS、MB-B仅在1:4时显示1级微弱的绿色荧光；MB在两个浓度梯度下均无荧光显示(0级).结论 携LIFR单抗通过生物素-亲和素桥连技术有效连接在MB-B微囊；体外荧光法是鉴定靶向脂质体微泡配体连接可靠性的简便方法.

Construction and fluorescence intensity of lipid ultrasound microbubbles with monoclonal antibody targeted to leukaemia inhibitory factor receptor

Objective To investigate fluorescence intensity of lipid ultrasound microbubbles constructed in vitro and targeted to leukaemia inhibitory factor receptor (LIFR) with a monoclonal antibody.Methods The LIFR-targeted ultrasound mierobubbles (MB-BSB-LIFR-AB) were constructed using a technology of biotin-avidin bridge.FITC labeled Avidin was incubated with lipid ultrasound microbubbles (MB) and biotinylated lipid microbubbles (MB-B).Two dilutions (1:4 and 1:16) of DTAF second antibody were incubated with four types of ultrasound microbubbles,including MB,MB-B,biotinavidin-MB (MB-BS),MB-BSB-LIFR-AB.The fluorescence intensity of microhubbles were graded as 0,1,2to 3.Results After incubating with FITC-avidin,MB-B displayed bright green fluorescence ( grade 3),but MB had no fluorescence ( grade 0).After incubating with two dilutions of DTAF second antibody (1:4 and 1:16),MB-BSB-LIFR-AB displayed brightest green fluorescence (grade 3) in both concentration,while MB-BS and MB-B only displayed dim green fluorescence (grade 1 ) at the dilution of 1:4,with MB displaying no fluorescence at either dilution (grade 0).Conclusions LIFR monoclonal antibody can be effectively conjugated to MB-B with biotin-avidin bridge.Fluorescence detection is a simple method for investigating the conjugation reliability of targeted lipid ultrasound microbubbles.