CAJ | 학술논문

目的构建携羊抗小鼠IgG单抗靶向超声造影剂(UCA-IgG)及体外评价其靶向黏附效能.方法采用"亲和素-生物素"桥接法构建UCA-IgG,体外荧光法鉴定IgG单抗与生物素化造影剂特异结合.利用平行板流动腔在不同时间点、不同浓度小鼠IgG包被和不同剪切应力下,测定相应的UCA-IgG的结合数目及达到半数解离时的剪切应力.设不同浓度小鼠包被的平行板流动腔为实验组,羊抗小鼠IgG封闭组和空白组为对照组.结果 UCA-IgG发出明亮的绿色荧光,而普通脂质造影剂无荧光显示.实验组UCA-IgG结合数量随包被浓度的增高而增加(P<0.05),并与结合时间呈明显正相关(P<0.05),然而与剪切应力呈现双向性(P<0.05).而两对照组均未见明显的UCA-IgG结合.UCA-IgG达半数解离的剪切应力随包被浓度的增加而增大(P<0.05).结论在不同血流动力学条件下UCA-IgG可与小鼠IgG特异有效结合,携羊抗小鼠IgG单抗造影剂可为其他特异靶向超声造影剂的体外研究提供有力的支持.
목적 구건휴양항소서IgG단항파향초성조영제(UCA-IgG)급체외평개기파향점부효능.방법 채용"친화소-생물소"교접법구건UCA-IgG,체외형광법감정IgG단항여생물소화조영제특이결합.이용평행판류동강재불동시간점、불동농도소서IgG포피화불동전절응력하,측정상응적UCA-IgG적결합수목급체도반수해리시적전절응력.설불동농도소서포피적평행판류동강위실험조,양항소서IgG봉폐조화공백조위대조조.결과 UCA-IgG발출명량적록색형광,이보통지질조영제무형광현시.실험조UCA-IgG결합수량수포피농도적증고이증가(P<0.05),병여결합시간정명현정상관(P<0.05),연이여전절응력정현쌍향성(P<0.05).이량대조조균미견명현적UCA-IgG결합.UCA-IgG체반수해리적전절응력수포피농도적증가이증대(P<0.05).결론 재불동혈류동역학조건하UCA-IgG가여소서IgG특이유효결합,휴양항소서IgG단항조영제가위기타특이파향초성조영제적체외연구제공유력적지지.
Objective To construct targeted ultrasound contrast agent carried goat anti-mouse IgG antibody (UCA-IgG) and evaluate the effectiveness of its targeted adhesion using parallel plate flow chamber. Methods The ultrasound contrast agent targeted to mouse IgG was designed by conjugating monoclonal antibodies against mouse lgG to the lipid monolayer shell of the agent using biotin-streptavidin. The binding of IgG antibodies to the ultrasound contrast agent were identified by fluorescence in vitro. The attachment and detachment of UCA-IgG to mouse IgG immobilized on a culture dish were assessed in a parallel-plate flow chamber. While the plate lacked mouse IgG,or blocked with large number of goat anti-mouse IgG were served as two control groups. Results UCA-IgG issued a bright green fluorescence, while the contral lipid ultrasound contrast agent didn't show fluorescence. The number of UCA-IgG bound to mouse IgG of experimental group was greater than two control groups,increased with increasing coverslips surface antibody concentrations (P<0. 05),and there was significant positive correlation between the number of UCA-IgG bound to mouse IgG and time of combination (P<0.05). The adhesion rate of experimental group increased with shear stress before 0. 5×10-5 N/cm2 (P<0.05) and then decreased (P<0. 05). There was limited adherence of control groups to the UCA-IgG. The stess of half-maximal detachment was increased with increasing coverslips surface antibody concentrations (P<0.05). Conclusions UCA-IgG could adhere to mouse IgG in the physical conditions. It may provide strong supports for studying other targeted ultrasound contrast agent preliminary and fatherly in vitro.