ActiveMax®靶点特异性激活磁珠：助力CAR细胞疗法高效富集与体外分析

文末福利来袭，别错过哦~

工程化免疫细胞疗法是肿瘤治疗领域的最新代表，尤其是嵌合抗原受体T细胞（CAR-T），在复发性或难治性多种血液系统癌症方面显示出显著疗效。由于CAR细胞是“活的药物”，其生产制备过程繁杂、质控要求严格，因此，迫切需要高质量的CAR细胞相关试剂/工具运用到药物开发中。研究显示，靶点抗原多聚物因其具有高靶点特异性、高精确性等特征在CAR细胞的富集分离、体外分析等多个场景广泛应用，尤其是基于磁珠偶联的特异性靶点多聚物展现出独特的应用优势。

• 关键应用2：精准助力CAR-T细胞的药代动力学（PK）研究，优化疗效监测

• 实验设计：

Flow plots showing prevalence of CD3+ CAR cells under magnetic enrichment with ActiveMax Human CD19 μbeads.

After positive selection with the ActiveMax Human CD19 ubeads at 2:4 (beads:ceels), the proportion of CAR cells that enriched cells were ≈150-fold higher than that of the initial population ~ 0.1%.

对CAR-T细胞的表型、在各种肿瘤微环境（包括不同抗原密度）中的激活数量和质量以及由此产生的效应功能进行精确且标准化的体外评估，对于成功开发CAR-T疗法并安全地应用于临床至关重要。在CAR-T细胞药物临床使用前的体外分析中，特异性靶点偶联磁珠可精确控制磁珠表面抗原密度，与CAR-T细胞共孵育后，通过流式细胞术或ELISA测定等手段，可以定量检测CAR-T细胞的抗原特异性、剂量依赖性激活等指标，有助于建立标准化的CAR-T细胞体外测定方法。

CAR-T细胞药物的效力（Potency）分析是批次放行（Lot Release Testing）的核心环节，直接影响产品的临床适用性。利用靶点特异性磁珠，可系统评估CAR-T细胞的激活能力及功能输出，确保每批次产品符合治疗标准。

在此，我们的ActiveMax® Human CD19 μBeads以2×10⁴ μBeads/孔（相当于约28.5ng偶联CD19蛋白）的密度接种于96孔板，并分别以效靶比（E:T）5:1，1:1，1:5与CD19 CAR-T细胞共培养24小时。单独培养的CD19 CAR-T细胞作为对照。随后，对活化标志物CD69、CD137进行检测分析，并对效应细胞因子IFN-γ和细胞毒性分子（颗粒酶B、穿孔素）进行激活后的分泌评估。结果显示，CD19 CAR-T细胞的CD69和CD137表达显著上调，证实了细胞的有效激活；刺激后的CAR-T细胞上清中IFN-γ分泌量明显增加，展现了强烈的效应功能。此外，ELISA检测还证实了颗粒酶B（Granzyme B）和穿孔素（Perforin）的分泌显著增强，表明ActiveMax® Human CD19 μBeads能有效诱导CAR-T细胞的细胞毒性作用。

• 实验设计：

CD19 CAR-T cells were stimulated with ActiveMax Human CD19 μBeads (MBS-C002) for 24h , and cells were harvested for evaluating CD69, CD137 expression by flow cytometry. The results showed that the proportion of CD69 and CD137 in CD19 CAR-T cells was significantly increased after CD19 μBeads stimulation.

CD19 CAR-T cells were stimulated with ActiveMax Human CD19 μBeads (MBS-C002) for 24h, and cell-free supernatants were harvested for evaluating IFN-γ secretion by flow cytometry. The results showed that CD19 CAR-T cells released significantly larger amounts of IFN-γ into the supernatants in response to CD19 μBeads.

CD19 CAR-T cells were stimulated with ActiveMax Human CD19 μBeads (MBS-C002) for 24h, and cell-free supernatants were harvested for evaluating IFN-γ secretion by ELISA. The results showed that CD19 CAR-T cells released significantly larger amounts of IFN-γ into the supernatants in response to CD19 μBeads.

• 实验结果三（细胞毒性分子分泌）：

CD19 CAR-T cells were stimulated with ActiveMax Human CD19 μBeads (MBS-C002) for 24h, and cell-free supernatants were harvested for evaluating Granzyme B, Perforin secretion by ELISA. The results showed that CD19 CAR-T cells released significantly larger amounts of Granzyme B, Perforin into the supernatants in response to CD19 μBeads.

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