Site-specific labeling of annexin V with F-18 for apoptosis imaging.
Site-specific labeling of annexin V with F-18 for apoptosis imaging.
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DOI：
10.1021/bc800164d
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发表时间：
2008-08
影响因子：
4.7
通讯作者：
Tait JF
中科院分区：
文献类型：
--
作者：
Li X;Link JM;Stekhova S;Yagle KJ;Smith C;Krohn KA;Tait JF
Annexin V通过结合暴露在细胞膜外表面的磷脂酰丝氨酸(PS)来检测凋亡细胞。在这项研究中，我们用硫醇选择性试剂18F-标记剂N-[4-[(4-[18F]fluorobenzylidene)aminooxy]butyl]maleimide([18F]FBABM)检测了Annexin V-128的标记，Annexin V是Annexin V的一种突变形式，在NH_2末端有一个半胱氨酸残基。我们还检测了18F标记的Annexin V-128([18F]Fan-128)的细胞结合亲和力。[18F]FBABM是在文献方法的基础上改进的两步一锅法合成。(TOYOKUNI等人，BioCondiate Chem.2003年，141253年−1259年)。[18F]FBABM的平均产率为23±4%(n=4，衰变校正)，比活度为6000Ci/∼。总合成时间为∼92min。这项研究的关键改进是鉴定并开发一种纯化方法来去除杂质N-[4-[(4-dimethylaminobenzylidene)aminooxy]butyl]maleimide 4，它的存在极大地降低了蛋白质标记的产率。[18F]FBABM与含有硫醇的膜联蛋白V-128偶联得到[18F]Fan-128，产率为37±9%(n=4，衰变校正)。[18F]Fan-128的红细胞结合实验表明，膜联蛋白V-128的这种修饰不影响其膜结合亲和力。因此，[18F]Fan-128作为细胞凋亡显像剂的体内研究是必要的。
Annexin V is useful in detecting apoptotic cells by binding to phosphatidylserine (PS) that is exposed on the outer surface of the cell membrane during apoptosis. In this study, we examined the labeling of annexin V-128, a mutated form of annexin V that has a single cysteine residue at the NH2 terminus, with the thiol-selective reagent 18F-labeling agent N-[4-[(4-[18F]fluorobenzylidene)aminooxy]butyl]maleimide ([18F]FBABM). We also examined the cell binding affinity of the 18F-labeled annexin V-128 ([18F]FAN-128). [18F]FBABM was synthesized in two-step, one-pot method modified from literature procedure. (Toyokuni et al., Bioconjugate Chem. 2003, 14, 1253−1259). The average yield of [18F]FBABM was 23 ± 4% (n = 4, decay-corrected) and the specific activity was ∼6000 Ci/mmol. The total synthesis time was ∼92 min. The critical improvement of this study was identifying and then developing a purification method to remove an impurity N-[4-[(4-dimethylaminobenzylidene)aminooxy]butyl]maleimide 4, whose presence dramatically decreased the yield of protein labeling. Conjugation of [18F]FBABM with the thiol-containing annexin V-128 gave [18F]FAN-128 in 37 ± 9% yield (n = 4, decay corrected). Erythrocyte binding assay of [18F]FAN-128 showed that this modification of annexin V-128 did not compromise its membrane binding affinity. Thus, an in vivo investigation of [18F]FAN-128 as an apoptosis imaging agent is warranted.
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影响因子：
3.1
作者：
Toretsky, J;Levenson, A;Mease, RC
通讯作者：
Mease, RC
DOI：
10.1016/j.ijrobp.2003.11.017
发表时间：
2004-07-01
影响因子：
7
作者：
Haas, RLM;de Jong, D;Verheij, M
通讯作者：
Verheij, M
影响因子：
3.1
作者：
Berndt, Mathias;Pietzsch, Jens;Wuest, Frank
通讯作者：
Wuest, Frank
影响因子：
2.9
作者：
Tait, JF;Gibson, DF;Smith, C
通讯作者：
Smith, C
DOI：
10.1007/s00259-003-1378-8
发表时间：
2004-04-01
影响因子：
9.1
作者：
Murakami, Y;Takamatsu, H;Nishimura, S
通讯作者：
Nishimura, S