Research Themes

• 質量分析法を用いた肥満による喘息の重症・難治化因子の同定と治療法の開発
  01 Apr. 2019 - 31 Mar. 2022
• Targeted steroidomics research for diagnosis of adrenocortical disease
  Grant-in-Aid for Scientific Research (C)
  01 Apr. 2013 - 31 Mar. 2016
  It is essentially pre-requisite for the prompt diagnosis and the treatment of diseases caused by congenital adrenocortical hyper-/hypoplasia originated from the chromosomal abnormality, because those disease are usually intractable. In this research, our purpose was directed for the development of the microdetermination of cortisol and its related precursors/metabolites in small amount of biological samples based on the use of proton-affinitive derivatization and LC-ESI-MS/MS. In addition, we investigated the syntheses of deuterated internal standards, 18-oxocortisol as an authentic sample and the preparation method of various kinds of proton-affinitive derivatives, and their utilities in LC-ESI-MS/MS. Of these derivatives, we revealed that the picolinate ester was the most useful, and the application of the present derivative for the diagnosis of diseases of the abnormality of adrenocortical hormone production was carried out.
• 前立腺がんの進展とアンドロゲンの動態解析　　　　　　　　　　　　　　　
  Apr. 2011 - Mar. 2012
• Development of the analytical method for estrogen-o-quinones and its application to the study on the initiation mechanism of human breast cancer
  Grant-in-Aid for Scientific Research (C)
  Tohoku Pharmaceutical University
  2010 - 2012
  Selective and sensitive analytical method for estrogen-o-quinones derived from catechol estrogen with enzymatic or non-enzymatic manner was developed based on the use of phenazine derivatization and LC-MS. The present method enabled to determine the catalytic activity of protein and some metal ions on the process of oxidation of catechol estrogens to the corresponding o-quinones. Furthermore, analytical method for the glutathione and L-cysteine conjugates of these o-quinones was established as one of the model for determining protein adducts of o-quinones.
• Development of ultrasensitive bioluminescent immunoassay by using DNA amplification technique
  Grant-in-Aid for Young Scientists (B)
  2009 - 2011
  In this study, a highly sensitive immunoassay was developed by using DNA amplification technique as detection system. Under isothermal condition, LAMP(Loop-mediated isothermal Amplification) and RCA(Rolling Circle Amplification) reaction enable successive DNA amplification. In order to apply DNA amplification to the detection system on immunoassay, DNA-conjugated antigen was prepared for competitive immunoassay. As a result, physically and chemically stable DNA provided highly efficient and sensitive signal amplification based on genetic engineering.
• Production of aptamer by SELEX technique and its application to diagnosis
  Grant-in-Aid for Scientific Research (C)
  Showa University
  2007 - 2009
  Aptamers are nucleic acid ligands that recognize a wide range of target molecules with high affinity and specificity. They are generated by in vitro selection of a randomized oligonucleotide pool in a technique known as systematic evolution of ligands by exponential enrichment (SELEX). The development of in vitro selection and amplification technology has allowed the identification of specific aptamers that bind target molecules with high affinity and even discriminate between closely related targets. Their binding affinities are comparable to those of antibodies and antigens. In comparison to antibodies, aptamers have the advantages of (i) simple preparation by chemical or enzymatic synthesis, (ii) thermal stability, (iii) discrimination of structural differences, (iv) ease of surface attachment for analysis, (v) ease of binding property modification through minor changes in sequence, (vi) minimization of experimental animal usage, and (viii) preparation for targets that are toxic or not inherently immunogenic. Aptamers are suitable as analytical, diagnostic, and therapeutic tools in applications requiring molecular recognition and are consequently gaining increased attention for their potential uses. In this study, to develop new diagnosis method, we developed sensitive and high performance aptamer binding assay using microchip electorophoresis, and prepared new aptamers for HMG (human menopausal gonadotropin), amyloid-β peptide and oxytocin. At first, a assay for Thronbin was developed using thrombin aptamer and microchip electrophoresis method, the result showed detection limit of 1 pmol. Next we prepared aptamer for HMG by SELEX, and HMG was detected by using the aptamer in a similar manner. In addition, amyloid-β peptide and oxytocin aptmaers were prepared.