| 학회 |
한국화학공학회 |
| 학술대회 |
2011년 봄 (04/27 ~ 04/29, 창원컨벤션센터) |
| 권호 |
17권 1호, p.348 |
| 발표분야 |
생물화공 |
| 제목 |
Enzyme Column with Trypsin Coated Polymer Nanofibers for Protein Digestion |
| 초록 |
We constructed a trypsin column for rapid and efficient protein digestion in proteomic analysis. Electrospun and ethanol-dispersed polymer nanofibers were used for the fabrication of highly stable trypsin coatings, which was prepared by a two-step process of covalent attachment and enzyme crosslinking. It has been observed that a simple step of ethanol dispersion improved not only the enzyme loading but also the performance of protein digestion. In-column digestion of enolase was successfully performed in less than twenty minutes. By applying the ethanol dispersion of polymer nanofibers, the bypass of samples was reduced by filling up the column with well-dispersed nanofibers, yielding more rigorous and reproducible digestions. Regardless of ethanol-dispersion or not, trypsin coatings showed better digestion performance and improved performance stability under recycled uses than covalently-attached trypsin, in-solution digestion, and commercial trypsin beads. The combination of highly stable trypsin coatings and ethanol-dispersion of polymer nanofibers has opened up a new potential to develop a trypsin column for on-line and automated protein digestion. |
| 저자 |
전승현1, 장문석1, 김병찬2, 이상원1, 김중배1
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| 소속 |
1고려대, 2한국과학기술(연) |
| 키워드 |
Enzyme coating; Enzyme column; Trypsin digestion; Enzyme stabilization
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| E-Mail |
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| 원문파일 |
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