抗D(IgM)血型定型试剂(单克隆抗体)国家参考品的研制

Preparation of national reference for anti-D(IgM)blood grouping reagent(monoclonal antibody)

目的研制抗D(IgM)血型定型试剂(单克隆抗体)国家参考品。方法将抗D(IgM)杂交瘤细胞培养上清液经0. 22μm滤膜无菌过滤分装,冻干制备抗D(IgM)血型定型试剂(单克隆抗体)国家参考品(简称参考品)。按YY/T 1238-2014《RhD(IgM)血型定型试剂(单克隆抗体)》行业标准,检测其外观、pH、特异性、亲和力及效价;并由4家实验室进行协作标定,进行瓶间精密度及热稳定性试验。结果参考品外观为白色冻干粉末,复溶后为无色或淡黄色透明液体,无摇不散的沉淀及异物;pH为7. 2;效价为1∶64;与RhD阳性红细胞有凝集反应,与RhD阴性红细胞无凝集反应;凝集时间均在15 s以内,3 min凝集块均大于1 mm2。经各实验室协作标定,随机抽取的2支参考品效价均为1∶64;37℃放置3、7、14 d的效价均为1∶64。结论成功制备抗D(IgM)血型定型试剂(单克隆抗体)国家参考品,其瓶间精密度及稳定性均较好,可用于抗D(IgM)血型定型试剂(单克隆抗体)效价检测的质量控制。

Objective To prepare the national reference for anti-D(IgM)blood grouping reagent(monoclonal antibody).Methods The culture supernatant of anti-D(IgM)hybridoma was filtered by using a 0.22μm filter membrane,filled and lyophilized to prepare the national reference for anti-D(IgM)blood grouping reagent(monoclonal antibody).According to the relevant requirements in Industry Standard for RhD(IgM)Blood Grouping Reagent(Monoclonal Antibody),the prepared national reference was tested for appearance,pH,specificity,affinity and titer,and was calibrated in collaboration by four laboratories to evaluate the precision between containers and thermal stability.Results The national reference was a white freeze-dried powder,which was colorless or light yellow transparent liquid without precipitate not dispersed on shaking or foreign matter after reconstitution.The pH value of the reference was 7.2,while the titer was1∶64.The reference showed agglutination with RhD positive cells while no agglutination with RhD negative cells.All the agglutinations appeared within 15 s,while the agglutination blocks formed in 3 min were more than 1 mm2 in size.Both the titers of two containers of reference in collaborative calibration were 1∶64.However,all the titers of references in stability test were 1∶64 after storage at 37℃for 3,7 and 14 d.Conclusion The national reference for anti-D(IgM)blood grouping reagent(monoclonal antibody)was successfully prepared,which showed high precision between containers and stability,and might be used for the determination of titer of anti-D(IgM)blood grouping reagent(monoclonal antibody).