National Institute of Biologicals, Ministry of Health and Family Welfare, Govt. of India, Sector 62, NOIDA, Uttar Pradesh, India.
National Institute of Biologicals, Ministry of Health and Family Welfare, Govt. of India, Sector 62, NOIDA, Uttar Pradesh, India.
Biologicals. 2020 May;65:33-38. doi: 10.1016/j.biologicals.2020.01.002. Epub 2020 Mar 6.
The assay of Anti T lymphocyte immunoglobulin for final drug product testing is carried out using flow cytometry on Peripheral Blood Mononuclear Cells (PBMCs) as specified in European and British Pharmacopeia. An alternate assay was developed wherein the potency based quality control evaluation of Anti T lymphocyte immunoglobulin is carried out by measuring complement dependent cytotoxicity (CDC) using fluorescent resazurin dye. The reported bioassay was specific, linear (R = 0.98), precise (%GCV for repeatability was 3.54% and intermediate precision was 4.27%) and accurate with relative bias of -5.54%. On the basis of results obtained from the repeated performances on single available product, system suitability criteria and sample acceptance criteria were proposed wherein Slope from 4 PL curve fit results for Reference Standard (RS) should be > 0.9, EC for RS should lie between 0.264 and 1.131 μg/ml and fold response should be > 2. Confidence interval range and estimated relative potency range obtained from the method validation were narrower than those mentioned for compendial method.
用于最终药物产品测试的抗 T 淋巴细胞免疫球蛋白检测采用流式细胞术，按照欧洲和英国药典中的规定，使用外周血单核细胞（PBMC）进行。开发了一种替代方法，其中通过使用荧光 Resazurin 染料测量补体依赖性细胞毒性（CDC）来进行基于效力的抗 T 淋巴细胞免疫球蛋白质量控制评估。报道的生物测定法具有特异性、线性（R=0.98）、精密度（重现性的%GCV 为 3.54%，中间精密度为 4.27%）和准确性，相对偏差为-5.54%。基于在单个可用产品上的重复性能获得的结果，提出了系统适用性标准和样品接受标准，其中 4-PL 曲线拟合结果的斜率应为参考标准（RS）>0.9，RS 的 EC 应介于 0.264 和 1.131μg/ml 之间，并且 fold response 应>2。方法验证获得的置信区间范围和估计的相对效力范围比药典方法中提到的更窄。
