单抗药物组织交叉反应中不同免疫组织化学方法的比较性研究

目的通过两种不同类型的单克隆抗体药物贝伐单抗和美罗华,探讨免疫组织化学常规二步法和直接法在人体组织交叉反应中的应用。方法免疫组织化学常规二步法采用相应二抗,通过底物酶显色,显示单克隆抗体药物与相应组织的结合情况。直接法采用标记后的单克隆抗体药物,通过生物素-链霉卵白素系统放大后显色,显示单克隆抗体药物与相应组织的结合情况。结果贝伐单抗采用直接法,其结果特异性和敏感性均高于常规二步法。美罗华直接法不能有效地显示该抗体与相应组织的结合,而常规二步法的结果特异性和敏感性均表现良好。结论不同类型单克隆抗体适合不同的免疫组织化学方法,最佳的实验方法是在尽可能减少背景性染色的同时,采用最低的浓度即能显示单克隆抗体与人体组织的特异性结合。

Comparative Study of Different Immunohistochemistry Methods in Cross-reactivity of Monoclonal Antibody Drugs

Objective To investigate the application of conventional two-step and one-step immunohistochemistry methods in tissue cross-reactivity through two different monoclonal antibody drugs(bevacizumab and rituximab).Methods It showed the monoclonal antibody binding to the target tissues by conventional two-step method(indirect immunohistochemistry method) choosing appropriate secondary antibody or one-step method(direct immunohistochemistry method) choosing biotinylated antibody.Results In bevacizumab study,the result of one-step method was better than that of two-step method in specificity and sensibility.On the contrary,in rituximab study,it showed rituximab binding to B lymphocyte by two-step method other than by one-step method.Conclusion The different immunohistochemistry methods were fit on the different monoclonal antibodies.The best experiment method was to display the specifical binding of monoclonal antibodies in the lowest concentration simultaneously to minimize background staining.