Objective: To detect the pyrogen in CAR-T cells product employing the HL60-IL-6 assay.

Methods: The HL60 cells were incubated with CAR-T cells injection or endotoxin standard for 48 hours. After then, the secreted cytokine interleukin-6 (IL-6) from HL60 cells was determined by ELISA. According to the four-parameter logistic curve fitted by Optical Density (OD) value corresponding to IL-6 and endotoxin standard concentration, the endotoxin equivalents of pyrogen content in the CAR-T cells products can be measured. Then, the method was validated, including the limit of detection (LOD), limit of quantitation, the recovery rate and the comparison of the determined results by HL60-IL-6 assay with that by the conventional pyrogen test, the Rabbit Pyrogen Test (RPT).

中文翻译：

---

HL60-IL-6检测在CAR-T细胞产物体外热原检测中的应用

目的：使用HL60-IL-6分析法检测CAR-T细胞产物中的热原。

方法：将HL60细胞与CAR-T细胞注射液或内毒素标准液孵育48小时。然后，通过ELISA测定HL60细胞分泌的细胞因子白介素-6（IL-6）。根据由对应于IL-6的光密度（OD）值和内毒素标准浓度拟合的四参数对数曲线，可以测量CAR-T细胞产物中热原含量的内毒素当量。然后，对该方法进行了验证，包括检测限（LOD），定量限，回收率以及HL60-IL-6测定与常规热原测试，兔热原测试（Rb）的测定结果的比较（ RPT）。