1996 Fiscal Year Final Research Report Summary
Development of rapid immmuno-assay methods utilizing HPLC and application to control of fermentation
| Project/Area Number |
07555561
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
生物・生体工学
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
KATOH Shigeo Kyoto University, Dept.Syn.& Biol.Chem.Associate Prof., 工学研究科, 助教授 (20026272)
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| Co-Investigator(Kenkyū-buntansha) |
MAJIMA Tsuyoshi NGK Insulators, Engineering Res., Chief Fellow, エンジニアリング事業本部, 主任研究員
TERASHIMA Masaaki Kyoto University, Dept.Syn.& Biol.Chem.Assistant Prof., 工学研究科, 助手 (30172092)
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| Project Period (FY) |
1995 – 1996
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| Keywords | Rapid immuno-assay / Control of fermentation / Sandwich method / Plont cell culture / Anti-peptide antibody / Specific elution |
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| Research Abstract |
For effective purification of useful bioproducts by genetically engineered microorganisms and cells, control and optimization of fermentation conditions are most important. Because measurable parameters are limited to temperature, pH and dissolved oxygen, it was impossible to control fermentation conditions depending on real-time measurements of the concentrations of target materials. In most cases the concentrations of bioproducts are determined by bioassay or enzyme linked immunosorbent assay methods, and these methods require half or one full day to get results. Therefore, a rapid assay method of bioproducts will be necessary for effective control of bioproduction systems.
In this work rapid immuno-assay methods utilizing HPLC columns and antibodies were developed and applied to control of fermentation. Performance of direct method and sandwich method using antibody-coupled HPLC columus were studied. By the former method it was possible to measure the concentration of bioproducts within 30min, if the effect of non-specific adsorption of impurities is minor. This method was applied to production of human alpha1-antitrypsin by culture of recombinant rice cells, in which a medium of a simple composition was used. The time-course of alpha1-antitrypsin production after induction was measured, and the suitable production period was determined.
A new sandwich method using anti-peptide antibody and peptide elution was developed and applied to fermentation systems contaning many impurities.
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