Development of a high-sensitive electrochemiluminescence immunosensor

高灵敏电化学发光免疫传感器的研制

• 批准号: 06650927
• 负责人: SAKAI Kiyotaka
• 金额: $ 1.34万
• 依托单位: Waseda University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06650927/
• 关键词: Electrochemiluminescence; Cathode; Luminol; Antibody-antigen reaction; Bovine plasma; Maleimide; Bonding selectivity; 抗体抗原反応; 量子収率; 反応次数; 抗体; 抗原; 過酸化水素; インジウムースズ酸化物

Antigens (human serum albumin (HSA), immunoglobulin G (IgG), human neutrophil lysozyme (Lys) were labeled with luminol using glutaraldehyde. Antigen-antibody reaction was performed, followed by H_2O_2 addition. Potential was applied to the indium-tin oxide (ITO) cathode in a flow-cell, and the electrochemiluminescence intensity was measured which was enhanced by the presence of antibody. Using this reaction, many antibodies may be determined. In bovine plasma, the intensity was lower than in water, because H_2O_2 concentration decreased due to the reaction with Fe ion. The intensity increased with antibody concentration, indicating that homogeneous antigen-antibody determination is possible. The same experiment was performed with antigens labled with luminol by the maleimide method. The electrochemiluminescence also increased with antibody concentration and higher antigen-antibody bonding selectivity was obtained which was not by the glutaraldehyde method. In bovine serum, the intensity fo both HSA and IgG was lower than in water. The intensity also increased with antibody concentration. Antibodies are also determinable by the maleimide method, if the luminescent reaction can be properly controlled using bovine serum. In conclusion, the novel antibody-determination method is proposed without washing using a flow cell.

抗原（人血清白蛋白（HSA），免疫球蛋白G（IgG），人类嗜中性粒细胞溶菌酶（Lys）使用卢甲醇标记使用戊二醛。进行抗原 - 抗体反应，然后添加H_2O_2。 ITO）在流动细胞中的阴极，并测量了使用该反应的抗体来增强的电化学强度。由于与Fe离子的反应，抗体浓度的强度增加，表明均匀的抗原 - 抗体测定是可以通过抗原方法进行的。 - 抗体键合的选择性是通过脂肪醛法获得的。强度也随抗体浓度增加。抗体也可以通过甲酰亚胺法确定，如果可以使用牛血清对发光反应进行适当控制。总之，提出了新型抗体确定方法，而无需使用流动池洗涤。

项目成果

Y.Yoshimi,N.Himi,K.Sakai: "Development of a continuous method of immunoassay with an electrochemiluminescence flow cell for monitoring antibody concentration antibody concentratio in body fluids" Better Living Through Innovative Biochemical Engineering (e

Y.Yoshimi、N.Himi、K.Sakai：“开发一种使用电化学发光流动池的连续免疫测定方法，用于监测体液中的抗体浓度抗体浓度”通过创新生化工程改善生活（e

Hirotaka Haramoto, Yasuo Yoshimi, Kiyotaka Sakai: "Development of ane electrochemiluminescent (ECL) immunosensor ; Determination of antibody concentration and electrochemiluminescence mechanism" Jpn J Artif Organs. 25(3). 683-687 (1996)

Hirotaka Haramoto、Yasuo Yoshimi、Kiyotaka Sakai：“电化学发光 (ECL) 免疫传感器的开发；抗体浓度的测定和电化学发光机制” Jpn J Artif Organs。

永見直之,吉見靖男,酒井清孝: "電気化学発光フローセルを用いた血中抗体センサの開発" 人工臓器. 23. 523-526 (1994)

Naoyuki Nagami、Yasuo Yoshimi、Kiyotaka Sakai：“利用电化学发光流动池开发血液抗体传感器”人工器官。 23. 523-526 (1994)

Y.Yoshimi,M.Himi,K.Sakai: "Development of a continuos method of immunoassay with an electrochemiluminescence flow cell for monitoring antibody concentration antibody concentration in body fluids" Better Living Through lnnovative Biochemical Engineering(ed

Y.Yoshimi、M.Himi、K.Sakai：“利用电化学发光流动池开发连续免疫测定方法，用于监测体液中的抗体浓度抗体浓度”通过创新生化工程改善生活（编辑）

Yasuo Yoshimi, Hirotaka Haramoto, Takehiro Miyasaka, Kiyotaka Sakai: "Cathodic electrochemiluminescence of luminol enhanced by antibody-antigen reaction" J.Chem.Eng.Japan. 29(5). 851-857 (1996)

Yasuo Yoshimi、Hirotaka Haramoto、Takehiro Miyasaka、Kiyotaka Sakai：“抗体-抗原反应增强鲁米诺的阴极电化学发光”J.Chem.Eng.Japan。