Development of super catalytic antibodies effective for allergy type I such as pollinosis

超催化抗体（Antigenase）是一种具有特异性结合抗原和酶促降解抗原双重功能的划时代分子。在本项目中，我们将生产用于治疗花粉症等I型变态反应的特征催化抗体。我们已经通过人IgE免疫Balb/c小鼠、细胞融合制备单克隆抗体、抗体DNA分析和分子模拟实验，揭示了某些单克隆抗体中存在催化三联体样结构。此外，我们研究了5 H5抗体对人IgE的降解活性。本研究对5 H5催化抗体的特性以及其他具有催化抗体能力的抗体进行了详细的研究。1、通过N-末端氨基酸序列分析确定了5 H5催化抗体对IgE的切割位点。该位点位于肥大细胞上IgE结合位点的上游。2、TP 41的降解速率（kcat） ...更多信息 3、对1 E3、4C 4和5 H10等单克隆抗体的重链和轻链cDNA序列进行分析。作为结果，假定三联体样结构存在于1 E3抗体的重链和轻链中。分离纯化单克隆抗体，为后续实验奠定基础。并以TP 41 -1多肽为底物，研究了其肽酶活性。观察到1 E3和5 H10抗体重链和轻链的活性。另一方面，未检测到4C 4重链的活性。4C 4轻链的肽酶活性尚不清楚。通过上述实验，除5 H5外，我们还获得了其他一些催化抗体。通过发现具有高kcat和低Km的催化抗体，我们将产生有效的催化抗体，用于治疗I型变态反应，如花粉症，特应性疾病等。

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 … More -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