Application of antibody engineering in forensic toxicological analysis

2014 Fiscal Year Final Research Report

• Project/Area Number: 24590859
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Legal medicine
• Research Institution: Kumamoto University
• Principal Investigator: SASAO Ako 熊本大学, 生命科学研究部, 助教 (80284751)
• Co-Investigator(Renkei-kenkyūsha): MORIOKA Hiroshi 熊本大学, 大学院生命科学研究部, 教授 (20230097) ; NISHITANI Yoko 熊本大学, 大学院生命科学研究部, 教授 (30359997) ; YONEMITSU Kosei 熊本大学, 大学院生命科学研究部, 准教授 (10128332)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: 薬毒物スクリーニング / 法医中毒学 / 組換え抗体 / 一本鎖抗体 / フルボキサミン / 向精神薬

Outline of Final Research Achievements

The aim of this study is to facilitate preparation of anti-drug antibodies　using antibody engineering such as phage display technology. In this study, we acquired cDNAs that encode immunoglobulin variable regions (VL and VH) from hybridoma cells producing anti-drug monoclonal antibodies (antidepressant, fluvoxamine; FLV, antipsychotic, phenothiazines; PTZ), and determined their DNA sequences. Then, we constructed expression vectors and prepared single chain variable fragment (scFv) recognizing FLV or PTZ. Their physicochemical properties were examined by ELISA or SPR analysis. Anti- FLV or anti-PTZ scFvs were found to have specific and sufficient binding to their each antigen. Also, we prepared an FLV Quenchbody protein that was a novel antibody-based fluorescent biosensor and examined its physicochemical properties.

Free Research Field

法医中毒学