Lentivirus capture directly from cell culture with Q-functionalised microcapillary film chromatography.
Lentivirus capture directly from cell culture with Q-functionalised microcapillary film chromatography.
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使用 Q 功能化微毛细管薄膜层析直接从细胞培养物中捕获慢病毒。
DOI：
10.1016/j.chroma.2012.06.072
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Darton NJ
中科院分区：
文献类型：
--
作者：
Darton NJ
基于塑料微毛细管膜（MCFs），开发了一种新型一次性吸附材料，用于纳米复合物的快速阴离子交换捕获，无需预过滤、澄清或样品预处理。使用乙烯-乙烯醇共聚物（EVOH）熔融挤出工艺制备包含19个平行微毛细管的MCF，每个微毛细管的平均内径为142±10μm。MCF内表面使用支链化学进行官能化以连接季胺基团，从而产生阴离子交换吸附剂。使用这种新制造的MCF-EV 0 H-Q的纳米复合物的纯化通过从预过滤的培养收获物中捕获慢病毒而被成功证明。发现该5 m层析基质结合并除去40%的结合慢病毒或2.5× 106感染单位（ifu）。通过从含有细胞和细胞碎片的未过滤未处理培养基中捕获慢病毒颗粒，进一步证明了该色谱基质允许大颗粒通过的独特性质。使用这种方法，56%或1× 107 ifu的捕获的慢病毒被洗脱。基于这种新材料的装置可能在临床慢病毒生产的早期阶段使用，直接从生物反应器中收获慢病毒颗粒。
A new disposable adsorbent material for fast anion-exchange capture of nano-complexes without prefiltering, clarification or pre-processing of samples was developed based on plastic microcapillary films (MCFs). An MCF containing 19 parallel microcapillaries, each with a mean internal diameter of 142±10μm, was prepared using a melt extrusion process from an ethylene–vinyl alcohol copolymer (EVOH). The MCF internal surfaces were functionalised using branched chain chemistries to attach quaternary amine groups producing an anion-exchange adsorbent. The purification of nano-complexes using this newly fabricated MCF–EVOH-Q was successfully demonstrated with the capture of lentivirus from pre-filtered culture harvest. This 5m chromatographic substrate was found to bind and elute ∼40% of bound lentivirus or 2.5×106infectious units (ifu). The unique properties of this chromatographic substrate that allow the passage of large particulates was further demonstrated with the capture of lentiviral particles from unfiltered un-processed culture media containing cells and cell debris. Using this approach, 56% or 1×107ifu of captured lentivirus was eluted. A device based on this new material might be used at an early stage in clinical lentiviral production to harvest lentiviral particles, directly from bioreactors.
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DOI：
10.1016/j.chroma.2011.01.030
发表时间：
2011-03
期刊：
Journal of chromatography. A
影响因子：
--
作者：
N. Darton;Nuno M. Reis;M. Mackley;Nigel K.H. Slater
通讯作者：
N. Darton;Nuno M. Reis;M. Mackley;Nigel K.H. Slater
影响因子：
5.4
作者：
Chan, L;Nesbeth, D;Darling, D
通讯作者：
Darling, D
影响因子：
12.4
作者：
Nesbeth, D;Williams, SL;Darling, D
通讯作者：
Darling, D
DOI：
10.1016/j.nano.2007.11.001
发表时间：
2008-03-01
影响因子：
5.4
作者：
Darton, Nicholas J.;Hallmark, Bart;Mackley, Malcolm R.
通讯作者：
Mackley, Malcolm R.