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为获得丰富的特异识别分子和实现无标记、高灵敏度检测,利用噬菌体展示技术,选择在pIII蛋白上展示12肽的噬菌体M13作探针,采用羧基-氨基共价耦联法制备噬菌体展示蛋白质芯片。在此基础上,与表面等离子体共振传感技术结合,使用Biacore3000仪器,检测噬菌体展示蛋白质芯片与特异性抗体的反应,测定反应动力学常数。实验结果表明:当抗体浓度为0.4μmol.L-1时,响应信号可达365±8个单位;不同浓度的抗体与芯片反应的数据较好地与S型曲线吻合。该方法可望用于噬菌体展示蛋白质芯片的检测。
In order to obtain a large number of specific recognition molecules and to realize label-free and high-sensitivity detection, phage display technology was used to select the phage M13 displaying the 12-peptide on the pIII protein as a probe. The phage display protein chip . On this basis, in combination with the surface plasmon resonance sensing technique, the Biacore 3000 instrument was used to detect the reaction of the phage display protein chip with the specific antibody and the reaction kinetic constant was measured. The results showed that when the concentration of antibody was 0.4μmol.L-1, the response signal was up to 365 ± 8 units. The data of different concentrations of antibody and chip reacted well with S-curve. This method is expected to be used for the detection of phage display protein chips.