Development of separation process for PEGylated positional isoforms based on the intrinsic interaction with specific structure on the solid-liquid interface
| Project/Area Number |
26820363
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Biofunction/Bioprocess
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| Research Institution | Yamaguchi University |
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Principal Investigator |
Yoshimoto Noriko 山口大学, 医学(系)研究科(研究院), 助教 (40432736)
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| Project Period (FY) |
2014-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2014: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
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| Keywords | PEG化タンパク質 / イオン交換クロマトグラフィー / 異性体分離 / chromatography / PEGylation / PEGylated protein / 変性 / リフォールディング |
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| Outline of Final Research Achievements |
The mechanism for the retention on ion exchange chromatography was analyzed with respect to the positional isoforms of PEGylated proteins by the liner salt salt gradient and pH gradient methods. The distribution coefficients of positional isoforms depended on pKa of modified lysine residue. The isoform eluted earlier when the modified lysine had low pKa values. The steric effect was found to be important in the separation of isoforms compared to their difference in charge on the basis of the retention behaviors of denatured proteins and PEGylated DNA. The diffusion coefficients of PEGylated proteins were determined in bulk solution and in the ion exchange resin to clarify the mass transfer properties in column.
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Report
(3 results)
Research Products
(8 results)
