【优势平台】Star Staining，助力CAR表达检测

对于CAR-T细胞来说，发挥肿瘤杀伤作用的有效成分是CAR阳性的T细胞，CAR-T细胞产品的包装规格及临床使用剂量是以CAR-T阳性细胞数表示的，因此，CAR转染阳性率是CAR-T细胞产品质量控制的必检项，监管部门建议应采用流式细胞法（FACS）检测CAR转染阳性率。荧光标记蛋白因其预先标记了荧光素基团，可通过一步染色完成检测，既简化了检测流程、节省了检测时间，又避免了因使用二抗产生的非特异背景，成为流式检测CAR表达的首选试剂。

然而，荧光标记蛋白的选择面临着诸多难题，比如使用传统随机化标技术导致标记后蛋白活性下降、荧光强度弱而无法检测低丰度表达的CAR、流式检测设备局限于可选择的荧光类型等问题。

ACROBiosystems凭借Star Staining新一代定点标记技术平台，成功开发了一系列荧光标记类型的CAR靶点相关蛋白产品，以“星级”的产品优势，全方位满足CAR-T阳性率检测的不同需求。

• 采用定点标记技术，高蛋白活性

• 高灵敏度和高特异性，经类CAR细胞批检验证

• 极低非特异信号，经PBMC双染批检放行

• 高度均一和批间一致性，高水平满足临床样本分析检测要求

• 高稳定性（经加速、冻融验证）

• 荧光类型丰富（FITCPEAPCAlexa Fluor 488555647）

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经MALS验证，Alexa Fluor 555-Labeled Human BCMA Star Staining产品纯度高于95%。

The purity of Alexa Fluor 555-Labeled Human BCMA / TNFRSF17 Protein, His Tag star staining (Cat. No. BCA-HA2H6) is more than 95% and the molecular weight of this protein is around 24-36 kDa verified by SEC-MALS.

Star Staining FITC-BCMA和PE-BCMA蛋白产品的检测灵敏度显著高于竞品。

Binding activity of FITC-Labeled Human BCMA and PE-Labeled Human BCMA protein from two different vendors were evaluated by the FACS analysis. The result showed that ACRO's Star Staining FITC-Labeled Human BCMA (Cat. No. BCA-HF2H3) and PE-Labeled Human BCMA (Cat. No. BCA-HP2H7) have a much higher binding activity than that of the other competitor.

Star Staining FITC-BCMA和PE-BCMA产品经未转染CAR的PBMC细胞双染批检测验证，无非特异背景问题，特异性显著高于竞品，保证CAR表达检测结果的特异性和准确性，适用于CAR-T临床样本分析。

Non-specific binding to non-transduced PBMCs between PE-Labeled Human BCMA Protein of Acro and competitor. 5e5 of non-transduced PBMCs were stained with PE -Labeled Human BCMA Protein and anti-CD3 antibody, washed and then analyzed with FACS. FITC signal was used to evaluate the expression of CD3+ T cells in non-transduced PBMCs, and PE signal was used to evaluate the non-specific binding activity to non-transduced PBMCs.

Non-specific binding to non-transduced PBMCs between FITC -Labeled Human BCMA Protein of Acro and competitor. 5e5 of non-transduced PBMCs were stained with FITC -Labeled Human BCMA Protein and anti-CD3 antibody, washed and then analyzed with FACS. PE signal was used to evaluate the expression of CD3+ T cells in non-transduced PBMCs, and FITC signal was used to evaluate the non-specific binding activity to non-transduced PBMCs.

Star Staining FITC-labeled Human BCMA产品可保持天然蛋白活性。

Binding activity of the Human BCMA before and after FITC labeling was evaluated in the above FACS analysis. The result shows that FITC-Labeled BCMA (Cat. No. BCA-HF2H3) and unconjugated Human BCMA have almost the same level of binding activity.

Binding activity of different lots of AF555-Labeled Human BCMA (Cat. No. BCA-HA2H6) and PE-Labeled Human BCMA (Cat. No. BCA-HP2H7) against anti-BCMA CAR-293 cells was evaluated by flow cytometry. The result shows very high batch-to-batch consistency.

FITC-Labeled Human BCMA (Cat. No. BCA-HF2H3) and PE-Labeled Human BCMA (Cat. No. BCA-HP2H7) at 25℃ for 48 hours and freeze-throw cycles are stable without performance reduction.