Chemically self-assembled antibody nanostructures as potential drug carriers.
Chemically self-assembled antibody nanostructures as potential drug carriers.
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DOI：
10.1021/mp300303k
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发表时间：
2012-11-05
影响因子：
4.9
通讯作者：
Wagner CR
中科院分区：
文献类型：
--
作者：
Fegan A;Kumarapperuma SC;Wagner CR
以双氢叶酸还原酶（DHFR）融合蛋白和双甲氨蝶呤（BisMTX）为原料，制备了具有单链可变片段（scFv）多拷贝的化学自组装抗体纳米结构（csan）。我们设计并合成了一种BisMTX化学二聚体（BisMTX - nh2），它含有第三个连接臂，可以与荧光团、放射性标签和药物结合。单价、二价和高阶antid3 CSANs与FITC标记的双甲氨蝶呤配体（bisMTX-FITC）组装，并被hhb - mlt（一种CD3+ t白血病细胞系）快速内化和运输到早期和晚期内核体和溶酶体。由于发现bisMTX-FITC掺入cans时的荧光明显大于游离配体，因此可以监测内吞噬的antid3 cans的稳定性。内化的csan在几个小时内是稳定的，而用无毒的DHFR抑制剂甲氧苄啶（TMP）治疗导致细胞荧光在几分钟内迅速丧失（约80%），这与纳米颗粒在细胞内的有效分解一致。在较长时间内（24小时），无论细胞是否用DMSO或甲氧苄氨嘧啶处理，细胞荧光下降75%-90%。虽然BisMTX是一种有效的DHFR抑制剂，但它对HPB-MLT细胞无毒（GI50 ~ 20uM）。相比之下，用BisMTX制备的antid3 CSANs在72小时的细胞毒性（GI50= 0.5-1.5 uM）比BisMTX高至少13倍。与我们在csan稳定性研究中的发现一致，使用甲氧苄啶治疗后未观察到细胞毒性增加。综上所述，我们的研究结果表明，用三功能BisMTX制备的细胞受体靶向csan可以作为潜在的组织选择性药物载体
Chemically self-assembled antibody nanorings (CSANs) displaying multiple copies of single chain variable fragments (scFv) can be prepared from dihydrofolate reductase (DHFR) fusion proteins and bismethotrexate (BisMTX). We have designed and synthesized a BisMTX chemical dimerizer (bisMTX-NH2) that contains a third linker arm that can be conjugated to fluorophores, radiolabels and drugs. Mono-, divalent and higher order AntiCD3 CSANs were assembled with a FITC labeled bis-methotrexate ligand (bisMTX-FITC) and found to undergo rapid internalization and trafficking by the HPB-MLT, a CD3+ T-leukemia cell line, to the early and late endosome and lysosome. Since the fluorescence of bisMTX-FITC when incorporated into CSANs was found to be significantly greater than the free ligand, the stability of the endocytosed AntiCD3 CSANs could be monitored. The internalized CSANs were found to be stable for several hours, while treatment with the non-toxic DHFR inhibitor trimethoprim (TMP) resulted in a rapid loss (>80%) of cellular fluorescence within minutes, consistent with efficient intracellular disassembly of the nanorings. Over longer time periods (24 h) cellular fluorescence decreased by 75%–90%, whether or not cells had been treated with DMSO or trimethoprim. Although BisMTX is a potent inhibitor of DHFR, it was found to be non-toxic (GI50 >20uM) to HPB-MLT cells. In contrast, AntiCD3 CSANs prepared with BisMTX were found to be at least 13- fold more cytotoxic (GI50=0.5–1.5 uM) than BisMTX at 72 hours. Consistent with our findings from CSANs stability studies, no increase in cytotoxicity was observed upon treatment with trimethoprim. Taken together, our results suggest that cell receptor targeting CSANs prepared with trifunctional BisMTX could be used as potential tissue selective drug carriers
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影响因子：
15
作者：
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通讯作者：
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5.2
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