Development of a high-sensitive electrochemiluminescence immunosensor

用鲁米诺和戊二醛标记抗原（人血清白蛋白（HSA）、免疫球蛋白G（IgG）、人中性粒细胞溶菌酶（Lys））。进行抗原抗体反应，然后加入H_2O_2。在流动池中向氧化铟锡（ITO）阴极施加电势，并测量电化学发光强度，该电化学发光强度由于抗体的存在而增强。使用该反应，可以测定许多抗体。在牛血浆中，由于H_2O_2与Fe ~（2+）发生反应，使H_2O_2浓度降低，因此其强度低于在水中的强度。强度随抗体浓度增加而增加，表明均匀抗原-抗体测定是可能的。用马来酰亚胺法用鲁米诺标记的抗原进行相同的实验。电化学发光强度也随抗体浓度的增加而增加，并获得了戊二醛法所没有的较高的抗原抗体结合选择性。在牛血清中，HSA和IgG的强度均低于水。强度也随抗体浓度增加而增加。如果使用牛血清可以适当控制发光反应，则也可以通过马来酰亚胺法测定抗体。总之，提出了一种新的抗体测定方法，无需使用流动池进行洗涤。

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.