Identification of Human scFvs Targeting Atherosclerotic Lesions
Identification of Human scFvs Targeting Atherosclerotic Lesions
复制标题
靶向动脉粥样硬化病变的人 scFv 的鉴定
DOI：
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发表时间：
2006
影响因子：
4.8
通讯作者：
G. Clofent
中科院分区：
文献类型：
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作者：
R. Robert;Marie;D. Daret;S. Miraux;A. Nurden;J. Franconi;G. Clofent
我们的目的是通过体内生物淘选研究动脉粥样硬化早期病变，并鉴定出病变区域的人类抗体。我们设计了一种两步法来快速分离人单克隆噬菌体展示单链抗体（MoPhabs），这些抗体与动脉粥样硬化动物模型中病变中发现的蛋白质反应。在单轮体内生物淘选后，从通过组织学和NMR显微镜鉴定的兔主动脉的患病切片洗脱MoPhabs。原位表达的MoPhab通过消减集落过滤筛选其识别动脉粥样硬化但不识别正常主动脉的能力来选择。通过我们的方法选择的MoPhab主要结合动脉粥样硬化病变。其中两个，B3.3G和B3.GER，作为scFv片段产生，识别存在于早期动脉粥样硬化病变表面和更复杂斑块内膜厚度内的表位。这些人MoPhab在体内注射后归巢于ApoE-/-小鼠中的动脉粥样硬化病变。亲和纯化了B3.3G识别的约56 kDa的蛋白，并通过质谱分析鉴定为玻连蛋白。这是第一次，单轮体内生物淘选已被用来选择人类抗体作为候选人的诊断成像和获得洞察显示在动脉粥样硬化斑块的目标。
Our aim was to investigate by in vivo biopanning the lesions developed early in atherosclerosis and identify human antibodies that home to diseased regions. We have designed a two-step approach for a rapid isolation of human Monoclonal phage-display single-chain antibodies (MoPhabs) reactive with proteins found in lesions developed in an animal model of atherosclerosis. After a single round of in vivo biopanning, the MoPhabs were eluted from diseased sections of rabbit aorta identified by histology and NMR microscopy. MoPhabs expressed in situ were selected by subtractive colony filter screening for their capacity to recognize atherosclerotic but not normal aorta. MoPhabs selected by our method predominantly bind atherosclerotic lesions. Two of them, B3.3G and B3.GER, produced as scFv fragments, recognized an epitope present on the surface in early atherosclerotic lesions and within the intimal thickness in more complex plaques. These human MoPhabs homed to atherosclerotic lesions in ApoE-/- mice after in vivo injection. A protein of ∼56 kDa recognized by B3.3G was affinity-purified and identified by mass spectrometry analysis as vitronectin. This is the first time that single round in vivo biopanning has been used to select human antibodies as candidates for diagnostic imaging and for obtaining insight into targets displayed in atherosclerotic plaques.
使用改良的集落提升选择程序从杂交瘤细胞系 T84.66 生产单链抗 CEA 抗体，以检测抗原阳性 ScFv 细菌克隆。
影响因子：
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作者：
Rodenburg,CM;Mernaugh,R;Bilbao,G;Khazaeli,MB
通讯作者：
Khazaeli,MB