Structural optimization of an aptamer generated from Ligand-Guided Selection (LIGS) resulted in high affinity variant toward mIgM expressed on Burkitt's lymphoma cell lines.
Structural optimization of an aptamer generated from Ligand-Guided Selection (LIGS) resulted in high affinity variant toward mIgM expressed on Burkitt's lymphoma cell lines.
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DOI：
10.1016/j.bbagen.2017.03.020
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发表时间：
2017-07
期刊：
影响因子：
--
通讯作者：
Mallikaratchy P
中科院分区：
文献类型：
--
作者：
Zümrüt HE;Batool S;Van N;George S;Bhandari S;Mallikaratchy P
适配体是一种合成的核酸短分子，能够识别特定的靶标。适体的选择使用一种筛选方法称为配体的系统进化指数富集（SELEX）。我们最近介绍了SELEX的一种变体，称为“配体引导选择”（LIGS），它允许识别针对已知细胞表面蛋白质的特定适体。利用LIGS，我们引入了三种针对膜结合IgM （mIgM）的特异性适配体，这是B细胞的标志。在选择的三个抗mIgM的适体中，被称为R1的适体被发现是有趣的，因为它能够识别靶细胞上的mIgM，然后阻断结合其抗原的抗igm抗体。我们系统地截断了亲本适配体R1，以设计具有增强亲和力的更短的变体。重要的是，本文中我们发现，最优化的R1适配体变体的特异性与抗igm抗体的特异性相似，这表明用于选择性洗脱适配体的配体的特异性决定了ligs生成的适配体的特异性。此外，我们报道了R1的截断变体能够在生理温度下识别migm阳性的人B淋巴瘤BJAB细胞，这表明ligs生成的适体可以重新优化为更高亲和力的变体。总的来说，这些发现表明了LIGS在产生高度特异性适配体方面的重要性，并具有潜在的生物医学应用前景。
Aptamers are synthetic, short nucleic acid molecules capable of specific target recognition. Aptamers are selected using a screening method termed Systematic Evolution of ligands by Exponential enrichment (SELEX). We recently have introduced a variant of SELEX called “LIgand-Guided-Selection” (LIGS) that allows the identification of specific aptamers against known cell-surface proteins. Utilizing LIGS, we introduced three specific aptamers against membrane-bound IgM (mIgM), which is the hallmark of B cells. Out of the three aptamers selected against mIgM, an aptamer termed R1, in particular, was found to be interesting due to its ability to recognize mIgM on target cells and then block anti-IgM antibodies binding their antigen. We systematically truncated parent aptamer R1 to design shorter variants with enhanced affinity. Importantly, herein we show that the specificity of the most optimized variant of R1 aptamer is similar to that of anti-IgM antibody, indicating that the specificity of the ligand utilized in selective elution of the aptamer determines the specificity of the LIGS-generated aptamer. Furthermore, we report that truncated variants of R1 able recognize mIgM-positive human B lymphoma BJAB cells at physiological temperature, demonstrating that LIGS-generated aptamers could be re-optimized into higher affinity variants. Collectively, these findings show the significance of LIGS in generating highly specific aptamers with potential applications in biomedicine.
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影响因子：
2.1
作者：
Maio G;Enweronye O;Zumrut HE;Batool S;Van N;Mallikaratchy P
通讯作者：
Mallikaratchy P
复杂靶标SELEX的演变以鉴定针对哺乳动物细胞表面抗原的适体。
DOI：
10.3390/molecules22020215
发表时间：
2017-01-30
期刊：
Molecules (Basel, Switzerland)
影响因子：
--
作者：
Mallikaratchy P
通讯作者：
Mallikaratchy P
影响因子：
15
作者：
Stojanovic, MN;Kolpashchikov, DM
通讯作者：
Kolpashchikov, DM
影响因子：
3.2
作者：
Shangguan, Dihua;Tang, Zhiwen;Tan, Weihong
通讯作者：
Tan, Weihong
影响因子：
--
作者：
EATON, BE;GOLD, L;ZICHI, DA
通讯作者：
ZICHI, DA