Developmental studies on practical use of ABO blood typing using immunoelectrodes

Research Project

Project/Area Number	03557030
Research Category	Grant-in-Aid for Developmental Scientific Research (B)
Allocation Type	Single-year Grants
Research Field	Legal medicine
Research Institution	Gunma University
Principal Investigator	FURUKAWA Ken Gunma Univ.Sch.Med.Dept.of Legal Med.Prof., 医学部, 教授 (90008227)
Co-Investigator(Kenkyū-buntansha)	NAKAJIMA Tamiko Gunma Univ.Sch.Med.Dept.of Legal Med.Assistant, 医学部, 助手 (40008561) MATSUKI Takasume Gunma Univ.Sch.Med.Dept.of Legal Med.Assistant Prof., 医学部, 助教授 (10126617) 矢沢伸群馬大学, 医学部, 講師 (10008386)
Project Period (FY)	1991 – 1993
Project Status	Completed (Fiscal Year 1993)
Budget Amount *help	¥4,600,000 (Direct Cost: ¥4,600,000) Fiscal Year 1993: ¥800,000 (Direct Cost: ¥800,000) Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000) Fiscal Year 1991: ¥2,600,000 (Direct Cost: ¥2,600,000)
Keywords	ABO blood groups / Blood group determination / Electrode / Membrane potential / Potential / 血液型 / 血液型物質 / 抗体 / 免疫電極 / 抗原抗体反応 / 免疫センサー / 型物質 / 化学修飾電極 / 銀塩化銀電極
Research Abstract	1) A titanium wire 1 mm in diameter was heated at ca.1000。C to make a thin film of oxide on the surface. The tip of the wire was immersed in a 2 mg/ml solution of cyanogen bromide pH11 for 20 min, washed and combined with anti-A, anti-B and eel anti-H antibody at pH 8.5 for 20 min. The electrode was sealed with Teflon. The reference electrode was prepared in the same manner except treatment with antibody. After the electrodes were set in the PBS with stirring, the corresponding human red cells or blood group substance was added to the PBS.A change in potential towards positive was observed after the addition of antigens. Addition of non-corresponding red cells or blood group substances did not change the potential. The result indicated that the ABO blood group determinations of red cells or blood group substances were possible using antibody fixed electrodes. 2) Human blood group stroma from blood group O, A and B red cell was fixed in a triacetyl-cellulose membrane. Electrochemical blood typing of membrane stroma was performed with antisera, using a pair of membranes with immobilized H, A or B stroma and stroma unfixed control membrane. The membrane-bound blood group substance retained its binding ability to the corresponding antibody in antiserum. The transmembrane potential changed after the antibody binding reactions with the membrane-fixed blood group substance. The blood group was determined measuring the membrane potential across these membranes before and after the antigen-antibody reaction.