| Budget Amount *help |
¥15,000,000 (Direct Cost: ¥15,000,000)
Fiscal Year 2006: ¥4,400,000 (Direct Cost: ¥4,400,000)
Fiscal Year 2005: ¥5,100,000 (Direct Cost: ¥5,100,000)
Fiscal Year 2004: ¥5,500,000 (Direct Cost: ¥5,500,000)
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| Research Abstract |
Super catalytic antibody (Antigenase) is an epochal molecule which can not only specifically bind to the antigens but also enzymatically degrade the antigen. In this project, we will produce characteristic catalytic antibodies for curing allergy type I such as pollinosis. We have already revealed the presence of catalytic triad-like structure in some monoclonal antibodies by the experiment of immunization of human IgE into balb/c mice, cell fusion for preparing monoclonal antibodies, DNA analysis of the antibodies, and molecular modeling. The, we investigated the degradation activity of 5H5 antibody against human IgE. In this year, we investigated the feature of the catalytic antibody in detail and other antibodies capable of being catalytic antibodies.
1, Cleaved sites of IgE by 5H5 catalytic antibody was determined by N-terminal amino acid sequencing analysis. The site was located at upstream of the binding site with which IgE can bind on mast cell.
2, The degrading rate (kcat) of TP41
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-1 peptide by the catalytic antibody 5H5 was 0.09/min.
3, CDNA sequence of the heavy and light chain of the monoclonal antibodies such as 1E3, 4C4 and 5H10 were analyzed. As the results, it was assumed that a triad-like structure was present in both heavy and light chain for 1E3 antibody. The structure was observed in the light chain.
4, The monoclonal antibodies were separated and purified for the following experiments. And the peptidase activity was investigated using TP41-1 peptide. The activity was observed for both heavy and light chain for 1E3 and 5H10 antibodies. On the other hand, no activity was detected for the heavy chain of 4C4. The peptidase activity was not clear for the light chain of 4C4.
Through the above experiments, we could obtain some other catalytic antibodies except for 5H5. By finding out the catalytic antibodies possessing high kcat and low Km, we will produce effective catalytic antibodies effective for curing the type I allergy such as pollinosis, atopic diseases etc. Less
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