| Project/Area Number |
12650756
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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 |
化学工学一般
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| Research Institution | Yamaguchi University |
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Principal Investigator |
YAMAMOTO Shuichi Yamaguchi University, Faculty of Engineering, Professor, 工学部, 教授 (80144921)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2000: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | biosensor / chromatography / immunoassay / antigen / antibody / Size-exclusion / ELISA / enzyme immuno assay / 抗原抗体複合体 / 免疫反応 / 抗体固定化 |
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| Research Abstract |
Various methods have been developed for immunoassay. The most popular method is enzyme-linked immunoadsorbent assay (ELISA), in which antibody is immobilized on the solid phase. In this study, free antigen-antibody complex was measured by using size exclusion chromatography (SEC). For comparison, factors affecting the precision and the reproducibility of 96-well microplate ELISA were examined experimentally. The items tested were antibody concentration, time for antibody immobilization, concentration of contaminant proteins, methods for washing, and etc.
Antibody and antigen-protein were successfully separated by micro-SEC (4.6 mm diameter and 30 cm length). However, it was found that quite large antibody-antigen complexes are formed during SEC separation even with monoclonal antibody. The present method is also suitable for characterizing monoclonal antibody properties.
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