| 研究課題/領域番号 |
20J15151
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| 研究機関 | 京都大学 |
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研究代表者 |
劉 晨晨 京都大学, 工学研究科, 特別研究員(DC2)
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| 研究期間 (年度) |
2020-04-24 – 2022-03-31
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| キーワード | poly(ethylene glycol) / electrophoresis / protein analysis / DNA analysis |
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| 研究実績の概要 |
For biomolecule separation, a new capillary electrophoresis (CE) method was developed by introducing a crosslinked bottle-brush polymer, copoly(poly(ethylene glycol) acrylate/poly(ethylene glycol) diacrylate) (copoly(PEGA/PEGDA)), into the capillary as a molecular sieving media. This study proved that the copoly(PEGA/PEGDA) gel can serve as a sieving matrix and afford comparable separation performance for 20-1500 bp DNA fragments to the authentic poly(acrylamide) (PA) gel. In addition, the copoly(PEGA/PEGDA) gel is able to yield a rapid and high-resolution separation at a proper PEGA/PEGDA monomer ratio. The compositions of the proposed hydrogel are commercially available and the synthetic protocol is similar to the commonly used PA gel, which facilitate the promotion of the present work.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
The proposed project involves the development of new hydrogel materials for the (1) molecular separation based on size sieving and (2) specific recognition based on the molecularly imprinted structure. Previously, we planned to synthesize the slide ring PEG hydrogel (Nat. Commun. 2014, 5, 5214) as the to-be-studied material. Now, we found the copoly(poly(ethylene glycol) acrylate/poly(ethylene glycol) diacrylate) (copoly(PEGA/PEGDA)) hydrogel shows a high separation performance as a sieving material and its feasibility in adjusting the molecular structure by a simple modification of the monomer ratio (ACS Appl. Polym. Mater. 2020, 2, 3886). Thus, the present project progresses with the copoly(PEGA/PEGDA) hydrogel instead of the slide ring PEG gel.
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| 今後の研究の推進方策 |
As for the molecular separation based on size sieving, a fundamental study is undergoing aiming at revealing the separation performance of copoly(PEGA/PEGDA)-hydrogel-based CE using standard protein markers as analytes.
For a specific recognition of proteins, we employ the molecularly imprinted (MP) technology to construct structural recognition sites towards target molecules. Here, a new method for fabricating the MP sites is proposed using structurally tailored and engineered macromolecular (STEM) gels (Macromolecules 2018, 51, 3808). Currently, we have established a general protocol to bring the 1st and 2nd reactions to reality. We are now working on using different proteins as templates to synthesize the MP sites via the two-step reaction.
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