3种抗人T淋巴细胞免疫球蛋白淋巴细胞毒效价检测方法比较  

Comparison of 3 determination methods for lymphocytotoxicity potency of anti-human T lymphocyte immunoglobulin

作　　者：杜天飞 于晓萍 李春阳[1] 刘兰军[1] Du Tianfei;Yu Xiaoping;Li Chunyang;Liu Lanjun(Recombinant Protein Drugs and New Vaccines Research Laboratory,Chengdu Institute of Biological Products Co.,Ltd,Center for Sichuan Provincial Vaccine Engineering Research,Chengdu 610023,China)

机构地区：[1]成都生物制品研究所有限责任公司重组蛋白质药物及新疫苗研究室,四川省疫苗工程技术研究中心,成都610023

出　　处：《国际生物制品学杂志》2024年第4期217-222,共6页International Journal of Biologicals

摘　　要：目的通过比较3种淋巴细胞毒效价检测方法测定抗人T淋巴细胞免疫球蛋白免疫血清及原液样品的效价结果, 阐明3种检测方法的相关性。方法采用中国药典2020年版三部淋巴细胞毒效价检测方法、基于人外周血单个核细胞的淋巴细胞毒流式检测方法及基于人白血病T细胞株(Jurkat细胞)的淋巴细胞毒流式检测方法测定3批次免疫血清及3批次原液的效价。免疫血清稀释128、256及512倍进行效价检测, 判定效价;原液稀释8~2 048倍进行检测, 计算半数效应浓度。结果中国药典方法与基于人外周血单个核细胞流式法检测3批次免疫血清的效价均为1∶512;基于Jurkat细胞流式法检测001批次免疫血清的效价为1∶512, 002与003批次免疫血清的效价为1∶256。3种方法测定3批次原液的效价结果与阳性对照Grafalon的活性水平相当, 不同方法检测的效价趋势基本一致。结论虽然3种方法测定免疫血清与原液的效价结果趋势基本一致, 但淋巴细胞毒流式检测方法操作更便捷, 且可定量测定半数效应浓度值。Objective To elucidate the correlation between three determination methods for lymphocytotoxicity potency of anti-human T lymphocyte immunoglobulin by comparing the titers of anti-human T lymphocyte immunoglobulin immune serum and bulk measured by 3 methods.Methods The titers of three batches of immune serum and bulk were determined by lymphocytotoxicity test in Chinese pharmacopoeia 2020 edition,peripheral blood mononuclear cell(PBMC)based and human leukemia T cell line(Jurkat cell)based lymphocyte cytotoxicity flow cytometry methods.The 128-,256-,and 512-fold diluted immune serum and the 8-to 2048-fold diluted bulk were tested for titer and median effective concentration(EC50).Results The titer results of immune serum determined by the method in Chinese pharmacopoeia and the PBMC based flow cytometry method were all 1:512.The titer of batch 001 immune serum determined by the Jurkat cell based flow cytometry method was 1:512,while the titers of batch 002 and 003 were 1∶256.The titers of three batches of bulk determined by the three methods were comparable to the positive control Grafalon,and the titer trends detected by different methods were basically consistent.Conclusion The trend of the three methods for determining the titers of immune serum and bulk is basically consistent,but the flow cytometry(PBMC and Jurkat)method is more convenient to operate and can quantitatively measure the EC50 value.

关 键 词：抗人T淋巴细胞免疫球蛋白 流式细胞术 淋巴细胞毒 效价 

分 类 号：R392-33[医药卫生—免疫学]