2020 Fiscal Year Annual Research Report
分子認識PEGゲルキャピラリー電気泳動による生体分子の構造・サイズ選択的分離
| Project/Area Number |
20J15151
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| Research Institution | Kyoto University |
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Principal Investigator |
劉 晨晨 京都大学, 工学研究科, 特別研究員(DC2)
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| Project Period (FY) |
2020-04-24 – 2022-03-31
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| Keywords | poly(ethylene glycol) / electrophoresis / protein analysis / DNA analysis |
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| Outline of Annual Research Achievements |
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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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
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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| Strategy for Future Research Activity |
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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