Viral vector purification with thermoresponsive-anionic mixed polymer brush modified beads-packed column
Viral vector purification with thermoresponsive-anionic mixed polymer brush modified beads-packed column
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使用热响应型阴离子混合聚合物刷修饰珠填充柱纯化病毒载体
DOI：
10.1016/j.seppur.2022.120445
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发表时间：
2022
影响因子：
8.6
通讯作者：
Kanazawa Hideko
中科院分区：
文献类型：
--
作者：
Nagase Kenichi;Kitazawa Sakiko;Kogure Toshihiko;Yamada Sota;Katayama Kazuhiro;Kanazawa Hideko
病毒载体作为基因治疗的潜在新治疗方式已引起人们的关注。在这项研究中，我们开发了一种温度调节的病毒载体纯化柱，使用由温敏和阴离子聚合物组成的混合聚合物刷作为填充材料配体。使用2-丙烯酰胺基-2-甲基丙磺酸（AMPS）的可逆加成-断裂链转移（RAFT）聚合和随后的N-异丙基丙烯酰胺（NIPAAm）的原子转移自由基聚合（ATRP）的组合在二氧化硅珠上修饰混合聚合物刷。温度调制zeta电位的变化，在所制备的混合聚合物刷归因于PNIPAAm收缩和暴露的PAMPS。以制备的混合聚合物刷修饰微球为包埋材料，观察了腺相关病毒2型（AAV 2）载体的洗脱行为。AAV 2载体在40 °C下通过静电和疏水相互作用吸附在混合聚合物刷上。通过将温度降低至5 °C，由于降低了AAV 2载体和混合聚合物刷之间的静电和疏水相互作用，混合聚合物刷上吸附的AAV 2载体被解吸并从柱洗脱。使用柱将AAV 2载体与作为污染物的牛血清蛋白分离。感染细胞的能力由从柱回收的AAV 2载体维持。因此，开发的柱将有利于病毒载体的简单纯化。
Viral vectors have attracted attention as a potential new therapeutic modality for gene therapy. In this study, we developed a temperature-modulated viral vector purification column using a mixed polymer brush composed of thermoresponsive and anionic polymers as packing material ligands. The mixed polymer brushes were modified on silica beads using a combination of reversible addition − fragmentation chain transfer (RAFT) polymerization of 2-acrylamido-2-methylpropanesulfonic acid (AMPS) and subsequent atom transfer radical polymerization (ATRP) ofN-isopropylacrylamide (NIPAAm). The temperature-modulated zeta potential change in the prepared mixed polymer brush was attributed to the PNIPAAm shrinking and exposing of PAMPS. The prepared mixed polymer brush modified beads were used as packing materials, and the elution behavior of the adeno associated virus type 2 (AAV2) vector was observed. The AAV2 vector was adsorbed on the mixed polymer brush by electrostatic and hydrophobic interactions at 40 °C. By reducing the temperature to 5 °C, adsorbed AAV2 vector on the mixed polymer brush was desorbed and eluted from the column due to lowering the electrostatic and hydrophobic interactions between the AAV2 vector and mixed polymer brush. The AAV2 vector was separated from bovine serum proteins as a contaminant using the column. The ability to infect cells was maintained by the recovered AAV2 vectors from the column. Thus, the developed column would be beneficial for the simple purification of viral vectors.
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影响因子：
4.3
作者：
Maekawa, Yutaro;Yamazaki, Kaichi;Kanazawa, Hideko
通讯作者：
Kanazawa, Hideko
智能阳离子热响应表面上腺苷核苷酸的温度调节相互作用变化1
DOI：
--
发表时间：
2007
期刊：
影响因子：
--
作者：
A. Kikuchi;J. Kobayashi;T. Okano;T. Iwasa;K. Sakai
通讯作者：
K. Sakai
影响因子：
5.4
作者：
Perabo, Luca;Goldnau, Daniela;Buening, Hildegard
通讯作者：
Buening, Hildegard
DOI：
10.1039/c7tb01251a
发表时间：
2017-08
期刊：
Journal of materials chemistry. B
影响因子：
--
作者：
K. Nagase;R. Shukuwa;T. Onuma;M. Yamato;N. Takeda;T. Okano
通讯作者：
K. Nagase;R. Shukuwa;T. Onuma;M. Yamato;N. Takeda;T. Okano
影响因子：
3.9
作者：
Nagase, Kenichi;Kobayashi, Jun;Okano, Teruo
通讯作者：
Okano, Teruo