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CAR-T靶点蛋白

荧光标记蛋白

CAR-T靶点蛋白

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生物素标记蛋白

生物素标记蛋白

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非标记蛋白

非标记蛋白

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嵌合抗原受体T（CAR-T）细胞疗法是一种基于体外改造T细胞，使其表面表达特异性识别肿瘤表面抗原的受体片段的癌症免疫疗法，改造后的T细胞输入患者体内，不需要抗原递呈细胞（APC）的帮助，直接靶向体内癌细胞并发挥免疫杀伤作用。

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质量控制的必检项，监管部门建议应采用流式细胞法检测CAR转染阳性率。目前有针对CAR不同结构区域的检测方法，包括针对CAR抗原结合位点的，比如CD19抗原，或针对轻链或铰链区的抗Fab抗体或Protein L蛋白，其中，针对抗原结合部位的CAR阳性率检测方法因其具有更好专属性而被广泛使用。

ACROBiosystems作为专注于医药研发领域的蛋白供应商，利用专业的蛋白研发平台、蛋白标记平台、稳定株开发平台和流式细胞分析平台，开发了一系列包括非标记、生物素标记、荧光标记等多种形式的CAR-T靶点蛋白以及配套的流式细胞法检测CAR阳性率的protocol，以助力CAR-T的研发，加速CAR-T研发的进程。目前产品已覆盖CD19、BCMA、CD22、MSLN和EGFR在内的近40个CAR-T热门靶点。

CAR-T阳性率检测策略及蛋白产品设计

直接检测法 —— 荧光标记蛋白

CAR-T靶点蛋白

产品特点

靶抗原预先标记了荧光素基团
可简化检测流程，节省检测时间
可避免因二抗交叉反应而引起的非特异性背景

> PE-labeled （点击分子即可查看产品详情）

BCMA Carbonic Anhydrase IX CD19 CD27 Ligand CD30 CD4 CD7 CD79B CLEC12A DLL3 EGF R EGFRvIII FAP FMC63 FOLR1 G4S linker Glypican 3 GUCY2C H-2Kb & B2M H-2Kb & B2M & OVA (SIINFEKL) H-2Kd & B2M Her2 HLA-A*0201 & B2M HLA-A*0201 & B2M & AFP (FMNKFIYEI) HLA-A*0201 & B2M & CMV pp65 (NLVPMVATV) HLA-A*0201 & B2M & EBV EBNA3C (LLDFVRFMGV) HLA-A*0201 & B2M & EBV LMP1 (YLLEMLWRL) HLA-A*0201 & B2M & EBV LMP2 (FLYALALLL) HLA-A*0201 & B2M & EBV LMP2A (CLGGLLTMV) HLA-A*0201 & B2M & HIV Gag (SLYNTVATL) HLA-A*0201 & B2M & hTERT (ILAKFLHWL) HLA-A*0201 & B2M & KRASG12V (KLVVVGAVGV) HLA-A*0201 & B2M & MAGE-A10 (GLYDGMEHL) HLA-A*0201 & B2M & p53 (HMTEVVRHC) HLA-A*0201 & B2M & PAP (ALDVYNGLL) HLA-A*0201 & B2M & Survivin (TLPPAWQPFL) HLA-A*0201 | B2M | HPV16-E6 HLA-A*0201 | B2M | HPV16-E7 HLA-A*0201 | B2M | MAGE-A4 (KVLEHVVRV) HLA-A*0201 | B2M | NY-ESO-1 HLA-A*0301 & B2M HLA-A*0301 & B2M & KRASG12V (VVGAVGVGK) HLA-A*1101 & B2M HLA-A*1101 & B2M & EBV (AVFDRKSDAK) HLA-A*1101 & B2M & EBV LMP2 (SSCSSCPLSK) HLA-A*1101 & B2M & HPV16-E6 (TTLEQQYNK) HLA-A*1101 & B2M & HPV16-E7 (IVCPICSQK) HLA-A*1101 & B2M & KRAS (VVVGAGGVGK) HLA-A*1101 & B2M & KRASG12D (VVVGADGVGK) HLA-A*1101 & B2M & KRASG12V (VVVGAVGVGK) HLA-A*2402 & B2M HLA-A*2402 & B2M & EBV EBNA3A (RYSIFFDYM) HLA-A*2402 & B2M & EBV EBNA3B (TYSAGIVQI) HLA-A*2402 & B2M & p53 (TYSPALNKMF) HLA-E*0103 & B2M HLA-E*0103 & B2M & CMV UL40 (VMAPRTLLL) HLA-E*0103 & B2M & CMV UL40 (VMAPRTVIL) HLA-E*0103 & B2M & CMV UL40 (VMAPRTVLL) HLA-E*0103 & B2M & CMV UL40 (VMPPRTVIL) IL-3 R alpha Mamu-A*01 & B2M Mesothelin Protein L PSMA ROR1 Siglec-2 Siglec-3 SIRP alpha SLAMF7 uPAR VEGF R2

> FITC-labeled （点击分子即可查看产品详情）

B7-H3 (4Ig) BCMA Carbonic Anhydrase IX CD19 CD27 Ligand CD30 CD37 CD38 CD4 CD47 CD5 CD7 CD72 CD8 alpha & beta CD98 CD99 CEACAM-5 CLEC12A DLL3 EGF R EGFRvIII EMMPRIN EpCAM ErbB3 FAP Flt-3 FMC63 FOLR1 Glypican 3 GUCY2C Her2 HGF R IL-13 R alpha 2 IL-3 R alpha LILRB4 Mesothelin NCAM-1 Nectin-4 NKG2D PSMA Rituximab ROBO1 ROR1 Siglec-2 Siglec-3 SLAMF7 Syndecan-1 uPAR VEGF R2

> 应用案例

使用FITC标记CD19蛋白检测Anti-CD19-CAR表达阳性率
: 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).; 点击申请Protocol

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基于生物素-亲和素的检测法 —— 生物素标记蛋白

生物素标记蛋白

产品特点

靶抗原预先标记了生物素，随后可使用荧光标记的链霉亲和素检测
链霉亲和素对生物素具有极高的亲和力，两者的结合具有高度特异性
可高灵敏度和高特异性地检测CAR阳性表达率

> Biotinylated proteins -> Specially designed

CA125 Carbonic Anhydrase IX CD19 CD30 CD38 CD4 CD5 CD7 CD72 CD8 alpha CD8 alpha & beta CEACAM-5 Chimeric HLA-A*0201 (H-2Kb α3) & B2M & GP100(YLEPGPVTA) CLEC12A DLL3 EGF R EGFRvIII EMMPRIN EpCAM EphA2 ErbB3 FAP FMC63 FOLR1 GUCY2C GUCY2D H-2Db & B2M H-2Kb & B2M & OVA (SIINFEKL) H-2Kb | B2M H-2Kd & B2M & InsB (LYLVCGERL) H-2Kd | B2M Her2 HLA-A*0101 & B2M HLA-A*0201 & B2M & CMV pp65 HLA-A*0201 & B2M & EBV EBNA (FMVFLQTHI) HLA-A*0201 & B2M & EBV EBNA3C (LLDFVRFMGV) HLA-A*0201 & B2M & EBV LMP1 (YLLEMLWRL) HLA-A*0201 & B2M & EBV LMP2 (FLYALALLL) HLA-A*0201 & B2M & EBV LMP2A (CLGGLLTMV) HLA-A*0201 & B2M & HER2 (KIFGSLAFL) HLA-A*0201 & B2M & HER2 (LIAHNQVRQV) HLA-A*0201 & B2M & HPV (TIHDIILECV) HLA-A*0201 & B2M & HPV16-E7 HLA-A*0201 & B2M & hTERT (ILAKFLHWL) HLA-A*0201 & B2M & KRAS (KLVVVGAGGV) HLA-A*0201 & B2M & KRASG12C (KLVVVGACGV) HLA-A*0201 & B2M & KRASG12D (KLVVVGADGV) HLA-A*0201 & B2M & KRASG12V (KLVVVGAVGV) HLA-A*0201 & B2M & KRASG12V (VVVGAVGVGK) HLA-A*0201 & B2M & MAGE-A10 (GLYDGMEHL) HLA-A*0201 & B2M & NY-ESO-1 (SLLMWITQV) HLA-A*0201 & B2M & p53 (HMTEVVRHC) HLA-A*0201 & B2M & p53 (HMTEVVRRC) HLA-A*0201 & B2M & PRAME (ALYVDSLFFL) HLA-A*0201 & B2M & PRAME (SLLQHLIGL) HLA-A*0201 & B2M & PSMA (MMNDQLMFL) HLA-A*0201 & B2M & SSX-2 (KASEKIFYV) HLA-A*0201 & B2M & Survivin (TLPPAWQPFL) HLA-A*0201 & B2M & Vaccinia virus (ILDDNLYKV) HLA-A*0201 & B2M & Vaccinia virus (ILYDNVVTL) HLA-A*0201 & B2M & Vaccinia virus (IMYDIINSV) HLA-A*0201 & B2M & Vaccinia virus (LLSYYVVYV) HLA-A*0201 & B2M & Vaccinia virus (SLSNLDFRL) HLA-A*0201 & B2M & Vaccinia virus (VLYNGVNYL) HLA-A*0201 & B2M & Vaccinia virus (YLAKLTALV) HLA-A*0201 | B2M & HIV Gag HLA-A*0201 | B2M & MAGE-A4 (KVLEHVVRV) HLA-A*0201 | B2M | AFP HLA-A*0201 | B2M | EW HLA-A*0201 | B2M | HBV HLA-A*0201 | B2M | MAGE-A1 HLA-A*0201 | B2M | MAGE-A2 HLA-A*0201 | B2M | MAGE-A3 HLA-A*0201 | B2M | MAGE-A4 HLA-A*0201 | B2M | p53 (GLAPPQHLIRV) HLA-A*0201 | B2M | p53 (RMPEAAPPV) HLA-A*0201 | B2M | RHAMM-R3 HLA-A*0206 & B2M HLA-A*02:01 & B2M HLA-A*02:01 & B2M & Glypican 3 HLA-A*02:01 & B2M & gp100 HLA-A*02:01 & B2M & NY-ESO-1 HLA-A*02:01 & B2M & WT-1 HLA-A*0301 & B2M HLA-A*0301 & B2M & gp100 (ALLAVGATK) HLA-A*0301 & B2M & KRAS (VVGAGGVGK) HLA-A*0301 & B2M & KRAS (VVVGAGGVGK) HLA-A*0301 & B2M & KRASG12C (VVGACGVGK) HLA-A*0301 & B2M & KRASG12C (VVVGACGVGK) HLA-A*0301 & B2M & KRASG12D (VVVGADGVGK) HLA-A*0301 & B2M & KRASG12V (VVGAVGVGK) HLA-A*0301 & B2M & Survivin Mut (RISTFKNWPK) HLA-A*1101 & B2M & EBV (AVFDRKSDAK) HLA-A*1101 & B2M & EBV LMP2 (SSCSSCPLSK) HLA-A*1101 & B2M & EGF R (KITDFGLAK) HLA-A*1101 & B2M & EGF R (KITDFGRAK) HLA-A*1101 & B2M & HPV16-E7 (IVCPICSQK) HLA-A*1101 & B2M & KRAS (VVVGAGGVGK) HLA-A*1101 & B2M & KRASG12A (VVVGAAGVGK) HLA-A*1101 & B2M & KRASG12C (VVVGACGVGK) HLA-A*1101 & B2M & KRASG12D (VVVGADGVGK) HLA-A*1101 & B2M & KRASG12S (VVVGASGVGK) HLA-A*1101 & B2M & KRASG12V (VVVGAVGVGK) HLA-A*1101 & B2M & MAGE-A1 (SLFRAVITK) HLA-A*1101 | B2M | KRAS (VVGAGGVGK) HLA-A*1101 | B2M | KRASG12D (VVGADGVGK) HLA-A*1101 | B2M | KRASG12V (VVGAVGVGK) HLA-A*11:01 & B2M HLA-A*2402 & B2M HLA-A*2402 & B2M & EBV EBNA3A (RYSIFFDYM) HLA-A*2402 & B2M & EBV EBNA3B (TYSAGIVQI) HLA-A*2402 & B2M & NY-ESO-1 (LLMWITQCF) HLA-A*2402 & B2M & p53 (TYSPALNKMF) HLA-A*2402 & B2M & Survivin-2B (AYACNTSTL) HLA-A*2402 & B2M & WT-1 (CYTWNQMNL) HLA-A*2601 & B2M HLA-A*3303 & B2M HLA-A*3303 & B2M & EGF R(HVKITDFGR) HLA-B*0702 & B2M HLA-B*0702 & B2M & HPV16 E7 (KPTLKEYVL) HLA-B*0801 & B2M HLA-B*1501 & B2M HLA-B*1502 & B2M HLA-B*2705 & B2M HLA-B*4001 & B2M HLA-B*5701 & B2M HLA-C*0303 | B2M HLA-C*0304 & B2M HLA-C*070201 & B2M HLA-C*0801 & B2M HLA-DRA1*0101 & HLA-DRB1*0401 HLA-DRA1*0101 & HLA-DRB1*1101 HLA-DRA1*0101 & HLA-DRB1*1101 & CLIP (PVSKMRMATPLLMQA) HLA-DRA1*0101 & HLA-DRB1*1101 & Influenza HA (PKYVKQNTLKLAT) HLA-DRA1*0101 & HLA-DRB1*1501 & MBP(ENPVVHFFKNIVTPR) HLA-E*0103 & B2M HLA-E*0103 & B2M & CMV UL40 (VMAPRTLLL) IL-13 R alpha 2 LILRB4 LY6G6D Mafa-A1*06301 & B2M & RM9 (RPKVPLRTM) Mamu-A*01 & B2M Mamu-A*08 & B2M Mesothelin NCAM-1 Nectin-4 NKG2D Protein L PSCA ROBO1 ROR1 Siglec-2 Siglec-3 SLAMF7 Syndecan-1 TRBC1 TRBC2 VEGF R2

> 应用案例

使用生物素标记BCMA蛋白检测Anti-BCMA-CAR表达阳性率
: 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); 点击申请Protocol

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间接检测法 —— 非标记蛋白

非标记蛋白

产品特点

靶抗原带有融合表位标签，随后可使用荧光标记的表位标签抗体作为二抗进行检测
可高灵敏度地检测CAR阳性表达率
有可能产生因二抗交叉反应引起的非特异性背景

> Unconjugated proteins -> Specially designed

BCMA CA125 CD171 CD19 CD1d&B2M CD30 CD38 CD38 & Serum Albumin CD4 CD5 CD7 CD72 CD8 alpha CD8 alpha & beta CEACAM-5 CLEC-2 Clec4b2 EGFRvIII EMMPRIN EpCAM EphA2 ErbB3 FAP FMC63 FOLR1 Glypican 3 gp350 GUCY2C Her2 HLA-A*0201 & B2M & NY-ESO-1 (SLLMWITQV) HLA-A*0201 & B2M & p53 (HMTEVVRRC) HLA-A*0201 & B2M & PRAME (SLLQHLIGL) HLA-A*0201 | B2M HLA-A*0201 | B2M | MAGE-A4 HLA-A*02:01 & B2M & Glypican 3 HLA-A*02:01 & B2M & MAGE-A4 (KVLEHVVRV) HLA-A*02:01 & B2M & NY-ESO-1 HLA-A*02:01 & B2M & WT-1 HLA-A*0301 & B2M HLA-A*0301 & B2M & KRAS (VVVGAGGVGK) HLA-A*0301 & B2M & KRASG12C (VVVGACGVGK) HLA-A*1101 & B2M HLA-A*1101 & B2M & KRAS (VVVGAGGVGK) HLA-A*1101 & B2M & KRASG12D (VVVGADGVGK) HLA-A*1101 & B2M & KRASG12V (VVVGAVGVGK) HLA-A*1101 | B2M | KRAS (VVGAGGVGK) HLA-A*1101 | B2M | KRASG12D (VVGADGVGK) HLA-A*1101 | B2M | KRASG12V (VVGAVGVGK) HLA-A*2402 & B2M HLA-A*2402 & B2M & EBV BRLF1 (TYPVLEEMF) HLA-DQA1*03:02 | DQB1*03:03 LY6G6D Mesothelin Mucin-1 NCAM-1 NKG2D Podoplanin Protein L ROBO1 ROR1 Siglec-2 Siglec-3 SLAMF7 Syndecan-1 TRAC TRBC1 TRBC2 TRDC TRGC1 TRGC2 uPAR VEGF R2

> 应用案例

使用Human CD19, Fc Tag蛋白检测Anti-CD19-CAR表达阳性率
: 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).; 点击申请Protocol

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更多CAR-T相关内容

> 点击查看更多Anti-FMC63 scFv Ab详细信息-更加灵敏检测Anti-CD19 (FMC63) CAR的特异性抗体 > 点击查看更多CD19详细信息——高亲和力CD19蛋白，专为检测CD19-CAR-T而设计

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