1996 Fiscal Year Final Research Report Summary
DEVELOPMENT OF COMPOSITE OF CHITOSAN AND DEXTRAN FOR SEPARATION OF PROTEINS AND ITS APPLICATION FOR INDUSTRIAL LARGE SCALE PROTEIN SEPARATION
| Project/Area Number |
07650926
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
反応・分離工学
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| Research Institution | Osaka Prefecture University |
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Principal Investigator |
YOSHIDA Hiroyuki Osaka Prefecture University Chemical Engineering Professor, 工学部, 教授 (50081360)
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| Co-Investigator(Kenkyū-buntansha) |
KONISHI Satoshi Osaka Prefecture University Chemical Engineering Research Associate, 工学部, 助手 (50275278)
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| Project Period (FY) |
1995 – 1996
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| Keywords | Chitosan / Dextran / Protein / Composite / Adsorption isotherm / Breakthrough curve / Chromatography / Compression |
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| Research Abstract |
Ion exchange chromatography is useful for large industrial-scale separation of proteins. A number of ion exchangers have been developed. Dextran-DEAE such as DEAE Sephadex A-50 (Pharmacia LKB Biotechnology) is a very good protein separator. However since it is very soft, it is compressed in a column even in very slow flow rate and therefore it can not be used in industrial-scale protein separation.
In the present work, we have developed a composite of chitosan and dextran-DEAE to make the dextran-DEAE hard. The supporter of the dextran-DEAE was a chitosan bead, Chitopearl 2501 (Ch-2501, Fuji Spinning Co.), which was a hard gel and an adsorber of protein in itself. We let dextran penetrate into the macropores of Ch-2501 by contacting with dextran solution. Thereafter, the dextran in the macropore was cross-linked and then DEAE was introduced on the dextran network. The finished product (hereafter called Ch-Dx-DEAE-2) was hard and was not compressed in a column at all. The equilibrium iso
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therm for adsorption of BSA was affected by pH considerably. This means that Ch-Dx-DEAE-2 is feasible for separation of proteins. The isotherms were correlated well by the Langmuir equation. The saturation capacity for adsorption of BSA on Ch-Dx-DEAE-2 was 4.5,2.5, and 1.3 times larger than those of DEAE Sepharose Fast Flow (Pharmacia LKB Biotechnology), Ch-2501, and DEAE Sephadex A-50, respectively. The experimental breakthrough curve agreed well with the theoretical breakthrough curve obtained by considering the intraparticle diffusion, liquid film diffusion, and axial dispersion in the bed. The intraparticle effective diffusivities of BSA were determined by matching the experimental breakthrough curves with the theoretical ones. The diffusivity in Ch-Dx-DEAE-2 was the largest in the above ion exchangers. It was ten times larger than that in Ch-2501. BSA adsorbed on Ch-Dx-DEAE-2 could be desorbed using NaCl aqueous solution easily. The elution curve of BSA from the bed was very sharp.The value of the peak was 870 times larger than that of feed concentration of BSA in adsorption process. This is 4 times larger than that obtained using the best commercial ion exchanger which is used commonly in the world. Less
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