2007 Fiscal Year Final Research Report Summary
Preparation of monoclonal antibody against lignin
| Project/Area Number |
17580143
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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 |
林産科学・木質工学
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| Research Institution | Kyoto University |
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Principal Investigator |
YOSHINAGA Arata Kyoto University, Graduate School of Agriculture, Assistant Professor (60273489)
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| Project Period (FY) |
2005 – 2007
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| Keywords | Tree cell wall / lignin / lignification / Immunolabeling / monoclonal antibody / 8-O-4 linkage / 8-5 linkage / 8-8 linkage |
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| Research Abstract |
To understand complex chemical structure of lignin in wood cell walls, monoclonal antibodies that recognize specific structure of lignin will be very important. The purpose of the present study is to develop the monoclonal antibody against lignin model compounds-protein conjugates. Three kinds of model compounds including three kinds of linkage (8-O-4, 8-5and8-8) of lignin were selected and conjugated with bovine serum albumin via p-aminohippuric acid. Monoclonal antibodies were raised with mice againstthree kinds of conjugates. As a result, 4hybridoma clones from 8-O-4 oligomer, 8 clones from 8-5 dimer and 8 clones from 8-8 dimer were developed.
Enzyme-linked immunosorbent assay showed that the monoclonal antibody against8-O-4 oligomer conjugates did not react with p-aminohippuric acid-BSA conjugates and that the antibody against 8-5 and 8-8 dimers did not react with both p-aminohippuric acid-BSA conjugates and 8-O-4 dimer (guaiacyl glycerol-beta-guaiacyl ether)-p-aminohippuric acid-BSA conjugates. These results suggested that these monoclonal antibodies could recognize different linkage structure of lignin.
Immunogold labeling of differentiating xylems of Japanese cypress, oak and poplar showed that the monoclonal antibody from at least one hybridoma clone from the three different antigens reacted with lignifying and lignified cell walls. Since the labeling sites were different among the three kinds of antigens, these monoclonal antibodies could recognize different chemical structure of lignin in wood cell walls. Although the specificity of these antibodies must be analyzed in detail, these monoclonal antibodies will be very effective probes to clarify complex structure of lignin and its formation process in wood cell walls.
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