压电免疫传感器抗体固化方法研究

Study on the methods for immobilization of piezoelectric immunobiosensor

推进了压电免疫传感器Ｃ_２型葡萄球菌肠毒素（ＳＥ）抗体三种固化方法（即：ＰＥＩ法、ＡＰＴＥＳ法、蛋白Ａ－金电极法）的研究，结果表明蛋白Ａ法固化抗体效果较好，其固化率达到８２．１３％。并对压电免疫传感器的性能进行了测试，其特异性、一致性经统计学分析，结果较好。抗体适宜浓度为５ｍｇ／ｍｌ。该传感器每次测过抗原后，用０．２ｍｏｌ／Ｌ的甘氨酸－盐酸洗脱后可重复使用３次，使用过的晶体片，用１．２ｍｏｌ／Ｌ的氢氧化钠、１．２ｍｏｌ／Ｌ的盐酸处理后，可循环使用多次。

Three different methods for immobilization of the C2 type staphylococcus enterotoxin (SE ) antibody on the gold electrode of the immunosensor were tested. Different methods resulted in some differences in terms of stability and sensitivity. Coating the electrode with protein A gave the best result ,immobilized ratio was 82. 13%. Specificity , consistency of piezoelectric immunobiosensor by statistic analysis presented better result. The best concentration of C2 type SE: antibody immobilized on electrode was 5mg/ml , the C2 type SE assay could be repeated for three times with the same crystal. For repeated uses , the adsorbed antigen could be removed from the crystal by washing the crystal with 0. 2mol/L Gly-HC1 without denaturing the immobilized antibody layer. Used quartz crystal were soaked in 1 . 2mol/L NaOH , then immersed in 1. 2mol/L HCl to remove all substances on the crystal surface ,and the quartz crystal can be used many times .