制备结合链酶亲和素超声造影剂的实验研究

杨莉; 刘政; 左松; 谭开彬; 高云华; 付赤学; 李秋颖

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中国临床医学影像杂志 ›› 2007, Vol. 18 ›› Issue (10) : 732-735.

论著

制备结合链酶亲和素超声造影剂的实验研究

杨莉1，刘政2，左松2，谭开彬2，高云华2，付赤学2，李秋颖2

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The preparation of streptoavidin conjugated ultrasound contrast agent: an experimental study

YANG Li1, LIU Zheng2, ZUO Song2, TAN Kai-bin2, GAO Yun-hua2, FU Chi-xue2, LI Qiu-ying2

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摘要

目的：探索制备一种结合链酶亲和素的脂膜超声微泡，以适用于亲和素－生物素法制备靶向超声造影剂，并评价其理化性质。方法：分4组于机械或超声振荡之前或之后，加入链酶亲和素完成超声造影剂的制备，采用浮选法洗涤。观察并检测洗涤前后各组微泡大小、形态、浓度、荧光亮度、微泡与链酶亲和素的结合情况。结果：机械或超声振荡制备的各组结合链酶亲和素的超声造影剂其浓度、形态与普通微泡比较无明显差异，但易静置分层。超声振荡法制备的微泡粒径稍大、浓度偏低。荧光显微镜观察：洗涤前各种不同方法所制备的微泡均能激发出明亮的红色荧光；洗涤后微泡浓度由1010左右降到107左右，微泡荧光亮度仍为“3级”。微泡与链酶亲和素的结合率，各种制备方法间比较差异无显著性意义（P>0.05），其结合率高达98%以上。结论：机械和超声振荡均可制备结合链酶亲和素的负电荷超声造影剂，微泡与其结合率高，为完成亲和素－生物素法制备靶向超声造影剂提供了重要基础。

Abstract

Objective: The purpose of this study is to prepare streptoavidin(SA) conjugated anionic lipid microbubbles and to assess the physiochemical properties. Methods: The first step was to direct binding of indocarbocyanin(Cy3) labeled SA to microbubbles by mechanical vibration, sonication, surface bind after mechanical vibration or sonication which were divided into four groups. Half of all resulted suspensions were used for repeated washings, trying to remove any free Cy3-SA using floatation method. Then, the size, shape, bubble concentration and fluorescent intensity were measured and analyzed under light and fluorescent microscopy. The binding rate of microbubbles with Cy3-SA was calculated using flow cytometry. Results: Compared with mechanical vibration groups, anionic lipid microbubbles with Cy3-SA in sonication groups appeared to be slightly larger in size. The microbubbles with Cy3-SA delaminated quicker than the control group. All Cy3-SA bubbles could give a bright red fluorescence except the control group. Though microbubbles concentration dropped with washings, it was still fluorescent visible and no significant binding rates differences were found in different preparation methods(P>0.05), it was above 98%. Conclusions: Streptoavidin can be conjugated with lipid microbubbles through mechanical vibration or sonication during preparation procedure. The conjugation of MB and SA were firmly high.

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造影剂 / 超声检查

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杨莉;刘政;左松;谭开彬;高云华;付赤学;李秋颖. 制备结合链酶亲和素超声造影剂的实验研究[J]. 中国临床医学影像杂志. 2007, 18(10): 732-735

YANG Li;LIU Zheng;ZUO Song;TAN Kai-bin;GAO Yun-hua;FU Chi-xue;LI Qiu-ying. The preparation of streptoavidin conjugated ultrasound contrast agent: an experimental study[J]. Journal of China Clinic Medical Imaging. 2007, 18(10): 732-735

中图分类号： Ｒ445.1