Controllable Molecular Sieving by copoly(Poly(ethylene glycol) Acrylate/Poly(ethylene glycol) Diacrylate)-Based Hydrogels via Capillary Electrophoresis for DNA Fragments
Controllable Molecular Sieving by copoly(Poly(ethylene glycol) Acrylate/Poly(ethylene glycol) Diacrylate)-Based Hydrogels via Capillary Electrophoresis for DNA Fragments
复制标题
通过基于共聚（聚（乙二醇）丙烯酸酯/聚（乙二醇）二丙烯酸酯）的水凝胶通过毛细管电泳进行 DNA 片段的可控分子筛分
DOI：
10.1021/acsapm.0c00567
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发表时间：
2020-09-11
影响因子：
5
通讯作者：
Otsuka, Koji
中科院分区：
文献类型：
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作者：
Liu, Chenchen;Kubo, Takuya;Otsuka, Koji
这项工作提出使用交联的共聚(聚(乙二醇)丙烯酸酯/聚(乙二醇)二丙烯酸酯)(共聚(PEGA/PEGDA))水凝胶作为毛细管电泳(CE)中DNA的筛分基质进行可控分子筛分。尝试使用不同的凝胶组分来控制分离性能，包括引发剂浓度（C-APS）、单体总浓度（C-t）、PEGA与PEGDA的比例（R）以及交联剂的长度（L-c）。结果表明，共聚(PEGA/PEGDA)凝胶中的分子筛分效应可以通过C-t、R、C-APS和L-c来控制，特别是前两个参数。较高的 C-t 和较低的 R 提供较小的平均孔径，从而更好地分离小尺寸 DNA 片段，而较低的 C-t 和较高的 R 则可以更好地分离大尺寸 DNA 片段。此外，使用代表性凝胶组分，重复测试显示迁移时间的逐次运行精度为 RSD < 0.9%（n = 10 次测试），柱间精度为 RSD < 5.9%（n = 5 根柱），日常精度为 RSD < 5.2%（n = 5 天内 25 次测试）。结果还表明，当调至较低的 R 和较低的 C-t 时，使用共聚 (PEGA/PEGDA) 凝胶可以快速、高分辨率地分离 20-140 bp DNA 片段。当使用高 C-t 时，发现共聚（PEGA/PEGDA）凝胶毛细管由于聚合时更好的体积稳定性而比真正的聚（丙烯酰胺）凝胶具有更好的重复性和更高的分辨率。
This work proposes controllable molecular sieving using the cross-linked copoly(poly(ethylene glycol) acrylate/poly(ethylene glycol) diacrylate) (copoly(PEGA/PEGDA)) hydrogels as a sieving matrix for DNA in capillary electrophoresis (CE). Attempts were made to control the separation performance using different gel components, including the initiator concentration (C-APS), total concentration (C-t) of monomers, the ratio (R) of PEGA to PEGDA, and the length of the cross-linker (L-c). Results reveal that the molecular sieving effect in the copoly(PEGA/PEGDA) gel can be controlled by C-t, R, C-APS, and L-c, especially by the former two parameters. Higher C-t and lower R provided smaller average pore size, resulting in better separation of small-size DNA fragments, while lower C-t and higher R allowed better separation of large-size DNA fragments. Moreover, using a representing gel component, the repeated test showed a run-to-run precision of RSD < 0.9% (n = 10 tests), a column-to-column precision of RSD < 5.9% (n = 5 columns), and a day-to-day precision of RSD < 5.2% (n = 25 tests in 5 days) for the migration time. The results also reveal the rapid and high-resolution separation for 20-140 bp DNA fragments using the copoly(PEGA/PEGDA) gel when tuned to a lower R and lower C-t. When high C-t was used, the copoly(PEGA/PEGDA) gel capillary was found to have better repeatability and higher resolution than the authentic poly(acrylamide) gels owing to the better volume stability upon polymerization.