Creation of the tailor-made system for antibody production by using an autoantigenic complex

本研究的目的是阐明核糖体自身抗原（P0-P1-P2复合物）的结构与抗原性之间的关系，并开发一种有用的抗体生产系统。核糖体自身抗原表位分析在c端发现了3个氨基酸，这是P0/P1/P2共享的，负责抗p识别。研究还发现，邻近3个氨基酸的丝氨酸残基磷酸化增强了抗p结合，表明自身抗原的磷酸化与抗原性有关。我们还发现，人类P1/P2的古细菌同源物aP1形成了一个稳定的四聚体，免疫该四聚体可产生针对保守c端部分的抗体。当将c端氨基酸序列替换为另一个核糖体蛋白序列时，产生针对引入序列的抗体。因此，我们开发了一种新的方法来有效地生产所需序列的抗体。

The aim of this study is to clarify the relationship between the structure and antigenicity of ribosomal autoantigen (P0-P1-P2 complex), and to develop a useful antibody-production system. Epitope analysis of the ribosomal autoantigen identified the 3 amino acids at the C-terminus, which is shared by P0/P1/P2 and responsible for anti-P recognition. It was also found that phosphorylation at Ser residues adjacent to the 3 amino acids enhanced the anti-P binding, suggesting that phosphorylation of the autoantigen is related to the antigenicity. We also found that aP1, the archaeal homologue of human P1/P2, forms a stable tetramer and that immunization of this tetramer resulted in production of antibodies to the conserved C-terminal part. When the C-terminal amino acid sequence was replaced with a sequence of another ribosomal protein, the antibody specific to the introduced sequence was produced. We thus developed a novel method to produce efficiently the antibody for a desired sequence.