EphB4-targeted imaging with antibody h131, h131-F(ab')2 and h131-Fab.
EphB4-targeted imaging with antibody h131, h131-F(ab')2 and h131-Fab.
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DOI：
10.1021/mp400354y
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发表时间：
2013-12-02
影响因子：
4.9
通讯作者：
Conti PS
中科院分区：
文献类型：
--
作者：
Li D;Liu S;Liu R;Zhou Y;Park R;Naga K;Krasnoperov V;Gill PS;Li Z;Shan H;Conti PS
越来越多的证据表明，酪氨酸激酶受体EphB 4的过度表达，血管发育的介质，是肿瘤诊断，预后和治疗的新靶点。因此，EphB 4表达的非侵入性成像对于在抗EphB 4治疗的最早阶段评估疾病过程和治疗功效可能是有价值的。在这项研究中，我们系统地研究了使用抗EphB 4抗体h131（150 kD）及其片段（h131-F（ab′）2，110 kD; h131-Fab，50 kD）用于近红外荧光（NIRF）成像的EphB 4体内表达。分别用胃蛋白酶和木瓜蛋白酶酶解h131，得到h131-F（ab′）2和h131-Fab，经FPLC和SDS-PAGE鉴定其纯度。在与Cy5.5缀合后，在EphB 4阳性HT 29肿瘤模型中评价h131、h131-F（ab′）2和h131-Fab的体内特征。虽然h131-Cy5.5在这些探针中表现出最高的肿瘤摄取，但其最佳肿瘤摄取水平在注射后2天（p.i.）获得。对于h131-Fab-Cy 5.5，在感染后4小时达到最大肿瘤摄取。而h131-Fab-Cy 5.5与hIgG-Fab-Cy 5. 5之间没有观察到显著差异，表明肿瘤积聚主要是由被动靶向引起的。相比之下，h131-F（ab′）2-Cy5.5在感染后6 h显示出显著的肿瘤摄取。通过hIgG-F（ab′）2-Cy5.5对照和免疫荧光染色证实靶特异性。总的来说，h131-F（ab′）2在早期时间点表现出显著和特异性的肿瘤摄取，这表明它是一种有前途的EphB 4靶向成像剂。
Accumulating evidence suggests that overexpression of the tyrosine kinase receptor EphB4, a mediator of vascular development, is a novel target for tumor diagnosis, prognosis and therapy. Noninvasive imaging of EphB4 expression could therefore be valuable for evaluating disease course and therapeutic efficacy at the earliest stages of anti-EphB4 treatment. In this study, we systematically investigated the use of anti-EphB4 antibody h131 (150 kD) and its fragments (h131-F(ab′)2, 110 kD; h131-Fab, 50 kD) for near-infrared fluorescence (NIRF) imaging of EphB4 expression in vivo. h131-F(ab′)2 and h131-Fab were produced through pepsin and papain digestion of h131 respectively, whose purity was confirmed by FPLC and SDS-PAGE. After conjugation with Cy5.5, in vivo characteristics of h131, h131-F(ab′)2 and h131-Fab were evaluated in EphB4-positive HT29 tumor model. Although h131-Cy5.5 demonstrated highest tumor uptake among these probes, its optimal tumor uptake level was obtained at 2 d post injection (p.i.). For h131-Fab-Cy5.5, maximum tumor uptake was achieved at 4 h p.i.. However, no significant difference was observed between h131-Fab-Cy5.5 and hIgG-Fab-Cy5.5, indicating the tumor accumulation was mainly caused by passive targeting. In contrast, h131-F(ab′)2-Cy5.5 demonstrated prominent tumor uptake at 6 h p.i. The target specificity was confirmed by hIgG-F(ab′)2-Cy5.5 control and immunofluorescent staining. Collectively, h131-F(ab′)2 exhibited prominent and specific tumor uptake at early time points, which suggests it is a promising agent for EphB4-targeted imaging.
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影响因子：
5.7
作者：
Spannuth WA;Mangala LS;Stone RL;Carroll AR;Nishimura M;Shahzad MM;Lee SJ;Moreno-Smith M;Nick AM;Liu R;Jennings NB;Lin YG;Merritt WM;Coleman RL;Vivas-Mejia PE;Zhou Y;Krasnoperov V;Lopez-Berestein G;Gill PS;Sood AK
通讯作者：
Sood AK
影响因子：
2.8
作者：
Wu, QH;Suo, ZH;Nesland, JM
通讯作者：
Nesland, JM
受体酪氨酸激酶EPHB4在卵巢癌中过表达，提供生存信号并预测结果不佳。
影响因子：
8.8
作者：
Kumar, S. R.;Masood, R.;Spannuth, W. A.;Singh, J.;Scehnet, J.;Kleiber, G.;Jennings, N.;Deavers, M.;Krasnoperov, V.;Dubeau, L.;Weaver, F. A.;Sood, A. K.;Gill, P. S.
通讯作者：
Gill, P. S.
影响因子：
4.9
作者：
Olafsen T;Wu AM
通讯作者：
Wu AM
DOI：
10.1006/bbrc.1996.1442
发表时间：
1996-09-24
影响因子：
3.1
作者：
Berclaz, G;Andres, AC;Crompton, MR
通讯作者：
Crompton, MR