嵌合抗原受體T（CAR-T）細胞療法是一種基于體外改造T細胞，使其表面表達特異性識別腫瘤表面抗原的受體片段的癌癥免疫療法，改造后的T細胞輸入患者體內，不需要抗原遞呈細胞（APC）的幫助，直接靶向體內癌細胞并發揮免疫殺傷作用。
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對于CAR-T細胞來說，發揮腫瘤殺傷作用的有效成分是CAR陽性的T細胞，CAR-T細胞產品的包裝規格及臨床使用劑量是以CAR-T陽性細胞數表示的，因此，CAR轉染陽性率是CAR-T質量控制的必檢項，監管部門建議應采用流式細胞法檢測CAR轉染陽性率。目前有針對CAR不同結構區域的檢測方法，包括針對CAR抗原結合位點的，比如CD19抗原，或針對輕鏈或鉸鏈區的抗Fab抗體或Protein L蛋白，其中，針對抗原結合部位的CAR陽性率檢測方法因其具有更好專屬性而被廣泛使用。
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ACROBiosystems作為專注于醫藥研發領域的蛋白供應商，利用專業的蛋白研發平臺、蛋白標記平臺、穩定株開發平臺和流式細胞分析平臺，開發了一系列包括非標記、生物素標記、熒光標記等多種形式的CAR-T靶點蛋白以及配套的流式細胞法檢測CAR陽性率的protocol，以助力CAR-T的研發，加速CAR-T研發的進程。目前產品已覆蓋BCMA，CD19，ROR1，EGFRVIII等20余個CAR-T熱門靶點.
CAR-T陽性率檢測策略及蛋白產品設計
直接檢測法 —— 熒光標記蛋白
產品特點
- 靶抗原預先標記了熒光素基團
- 可簡化檢測流程，節省檢測時間
- 可避免因二抗交叉反應而引起的非特異性背景
> FITC-labeled （點擊分子即可查看產品詳情）-> 獨有產品
| BCMA | CD19 | CD22 | GPC3 | MSLN | Protein L |
> PE-labeled （點擊分子即可查看產品詳情）-> 獨有產品
| MSLN |
> 應用案例
使用FITC標記CD19蛋白檢測Anti-CD19-CAR表達陽性率
293 cells were transfected with anti-CD19-scFv and RFP tag. 2e5 of the cells were stained with B. FITC-labeled Human CD19 (20-291) (Cat. No. CD9-HF2H2, 10 μg/ml) and C. FITC-labeled protein control. A. Non-transfected 293 cells and C. FITC-labeled protein control were used as negative control. RFP was used to evaluate CAR (anti-CD19-scFv) expression and FITC was used to evaluate the binding activity of FITC-labeled Human CD19 (20-291) (Cat. No. CD9-HF2H2).
> 如需了解更多此產品信息，可點擊：ACROBiosystems官網，輸入貨號，即可查看產品詳情，也可直接撥打：400-682-2521進行咨詢
基于生物素-親和素的檢測法 —— 生物素標記蛋白
產品特點
- 靶抗原預先標記了生物素，隨后可使用熒光標記的鏈霉親和素檢測
- 鏈霉親和素對生物素具有極高的親和力，兩者的結合具有高度特異性
- 可高靈敏度和高特異性地檢測CAR陽性表達率
> AvitagTM biotinylated proteins -> Specially designed
| BCMA | CD22 | CD30 | CD33 | CD38 | CD70 |
| EGF R | EGFRVIII | EpCAM | FOLR1 | GPC3 | HER2 |
| HGFR | MSLN | ROR1 |
> Chemically biotinylated proteins -> Specially designed
| CD19 | EpCAM | HER2 | MSLN | Protein L |
> 應用案例
使用生物素標記BCMA蛋白檢測Anti-BCMA-CAR表達陽性率
Human T cells were transfected with anti-BCMA CAR and cultured for 3 days. Three days post-transfection, 1e6 cells were first incubated with 50 μl biotinylated human BCMA protein (Cat. No. BC7-H82F0, 8 μg/ml), washed and then stained with PE Streptavidin and analyzed by flow cytometry. (Data are kindly provided by PREGENE Biopharma)
> 如需了解更多此產品信息，可點擊：ACROBiosystems官網，輸入貨號，即可查看產品詳情，也可直接撥打：400-682-2521進行咨詢
間接檢測法 —— 非標記蛋白
產品特點
- 靶抗原帶有融合表位標簽，隨后可使用熒光標記的表位標簽抗體作為二抗進行檢測
- 可高靈敏度地檢測CAR陽性表達率
- 有可能產生因二抗交叉反應引起的非特異性背景
> Fc-tag fusion proteins -> Specially designed
| BCMA | CD19 | CD30 | CD33 |
| CD38 | CD70 | EGFR | FOLR1 |
| GPC3 | HER2 | HGFR | IL13R |
| MSLN | MUC1 | ROR1 | EpCAM |
> His-tag fusion proteins -> Specially designed
| BCMA | CD19 | CD123 | CD138 | CD22 | CD30 |
| CD33 | CD38 | CD70 | CAIX | EGFR | EGFRVIII |
| EpCAM | FOLR1 | GPC3 | HER2 | HGFR | MSLN |
| ROR1 |
> All fusion proteins -> Specially designed
| BCMA | CD19 | CD123 | CD138 | CD22 | CD30 |
| CD33 | CD38 | CD70 | CAIX | EGFR | EGFRVIII |
| EpCAM | FOLR1 | GPC3 | HER2 | HGFR | IL13RA2 |
| MSLN | MUCI | ROR1 |
> 應用案例
使用Human CD19, Fc Tag蛋白檢測Anti-CD19-CAR表達陽性率
293 cells were transfected with FMC63-scFv and RFP tag. 2e5 of the cells were first stained with B. Human CD19 (20-291) Protein, Fc Tag, low endotoxin (Super affinity) (Cat. No. CD9-H5251, 3 μg/ml) and C. Human Fc Tag Protein Control, followed by FITC-conjugated Anti-human IgG Fc Antibody. A. Non-transfected 293 cells and C. Human Fc Tag Protein Control were used as negative control. RFP was used to evaluate CAR (anti-CD19-scFv) expression and FITC was used to evaluate the binding activity of Human CD19 (20-291) Protein, Fc Tag, low endotoxin (Super affinity) (Cat. No. CD9-H5251).
> 如需了解更多此產品信息，可點擊：ACROBiosystems官網，輸入貨號，即可查看產品詳情，也可直接撥打：400-682-2521進行咨詢
更多CAR-T相關內容
> 點擊查看更多CD19詳細信息——高親和力CD19蛋白，專為檢測CD19-CAR-T而設計
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