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"膜杰作"—多次跨膜靶点蛋白开发技术平台及系列产品
背景
图1．多次跨膜靶点蛋白（ACROBiosystems均可提供）跨膜蛋白是嵌入细胞膜磷脂双分子层中实现细胞内外跨越的一类蛋白，跨膜区域的相互作用是连接膜外环境与细胞内环境的重要渠道，以此来执行各种激活和应答反应实现信号转导，调控细胞内外的形态和功能上的改变。许多人类疾病或许都与异常的跨膜蛋白功能有关，跨膜蛋白的多功能性使它们成为理想的药物作用靶点。具有全长完整结构的多次跨膜靶点蛋白具有重要意义，以CD20为例，已有研究证实Rituximab对CD20的识别不止有一级表位ECL2，还识别其二级表位ECL1/ECL2，二级表位ECL1/ECL2还会影响Rituximab的CDC效应[1]。然而，多次跨膜蛋白结构复杂，具有多个疏水跨膜区，往往在宿主细胞中表达水平极低。针对多次跨膜靶点蛋白全长的制备难点，为助力多次跨膜蛋白靶标的药物研究，ACROBiosystems专门搭建了多次跨膜蛋白研发生产的更全面的平台化解决方案，满足跨膜靶点蛋白不同应用场景的需求，提供种类齐全的全长多次跨膜蛋白，包括四次跨膜蛋白CD20、Claudin18.2、Claudin 6，五次跨膜蛋白CD133，七次跨膜蛋白GPRC5D、CXCR4、CCR5、CCR8（图1）。三大技术平台技术原理ACROBiosystems专门搭建了基于HEK293表达系统的VLP技术平台，将构象完整的膜蛋白直接聚集在细胞表面，使这些复杂的膜蛋白转变为可溶性的、高浓度的蛋白用于抗体免疫和筛选，所制备的包膜VLPs在其固有的细胞膜上显示正确折叠的多次跨膜蛋白，从而能够诱导和筛选识别靶标天然构象的功能性抗体。
ACROBiosystems除提供VLP技术平台下常规生产的多次跨膜蛋白之外，同时也接收 定制服务。独特优势全长跨膜蛋白
VLP中靶抗原丰度高于过表达细胞
可提高免疫原性
100-300 nm，可作为树突状细胞和噬菌体展示的最佳靶点
可用于免疫/ELISA/SPR/BLI/细胞实验/CAR阳性率检测技术原理多次跨膜蛋白的跨膜区域疏水性较高，在普通的缓冲液中很难保持正确的构象，可以通过添加去垢剂解决该问题。以昆虫细胞表达技术和哺乳动物细胞表达技术为基础，ACROBiosystems专门搭建了困难药物靶点跨膜蛋白的表达平台和纯化平台，并且已经从常用的去垢剂中筛选出包括DDM/CHS（货号DC-11）在内的几种效果很好的去垢剂，大大增加多次跨膜靶点蛋白的水溶性，帮助其在溶液中形成天然正确的构象。
独特优势具有天然完整构象
可精确定量
可应用于免疫/ELISA/SPR/BLI技术原理“Nanodisc”的磷脂层结构，是由膜支架蛋白(membrane scaffold proteins, MSPs)和磷脂分子构成的磷脂双分子层类膜结构，跨膜蛋白移除去垢剂后可以整合到Nanodisc的特殊结构中，保持其生物学活性并有很好的水溶性，应用更广泛。ACROBiosystems研发团队经过不断优化改进，已成功摸索出适用于工业化放大生产的Nanodisc组装工艺，实现了跨膜蛋白Nanodisc产品的长期稳定供货。
独特优势全长的跨膜蛋白处于近似天然膜环境中，保持生物活性
具有良好的水溶性，无需去垢剂
可用于免疫/ELISA/SPR/BLI/细胞实验/CAR阳性率检测产品列表- VLP平台
- 膜蛋白-去垢剂平台
- Nanodisc平台
分子 货号 产品描述 应用 订购/预购 ADORA2A ADA-H52P4 Human ADORA2A Full Length Protein (VLP) 免疫
ELISA
SPR/BLI
细胞实验
CAR阳性率检测CCR5 CC5-H52P3 Human CCR5 Full Length Protein (VLP) CCR8 CC8-H52P4 Human CCR8 Full Length Protein (VLP) CD133 CD3-H52P3 Human CD133 Full Length Protein (VLP) CD151 CD1-H52P9 Human CD151 Full Length Protein (VLP) CD20 CDP-H52P6 Human CD20 Full Length Protein (VLP) CD20 CDP-HF2P8 Fluorescent Human CD20 Full Length Protein (VLP) CD37 CD7-H52P4 Human CD37 Full Length Protein (VLP) CD81 CD1-H52P5 Human CD81 Full Length Protein (VLP) Claudin-1 CL1-H52P6 Human Claudin-1 Full Length Protein (VLP) Claudin-18.1 CL1-H52P3 Human Claudin-18.1 Full Length Protein (VLP) Claudin-18.2 CL2-H52P7 Human Claudin-18.2 Full Length Protein (VLP) Claudin-18.2 CL2-HF218 Fluorescent Human Claudin-18.2 Full Length Protein (VLP) Claudin-4 CL4-H52P5 Human Claudin-4 Full Length Protein (VLP) Claudin-6 CL6-H52P7 Human Claudin-6 / CLDN6 Full Length Protein (VLP) Claudin-6 CL6-HF2G8 Fluorescent Human Claudin-6 / CLDN6 Full Length Protein (VLP) Claudin-9 CL9-H52P5 Human Claudin-9 / CLDN9 Full Length Protein (VLP) CNR1 CN1-H52P6 Human CNR1 Full Length Protein (VLP) CXCR3 CX3-H52P4 Human CXCR3 Full Length Protein (VLP) CXCR3 CX3-M52P7 Mouse CXCR3 Full Length Protein (VLP) CXCR4 CX4-H5219 Human CXCR4 / CD184 Full Length Protein (VLP) CXCR5 CX5-H52P4 Human CXCR5 Full Length Protein (VLP) CXCR6 CX6-H52P3 Human CXCR6 / CD186 Full Length Protein (VLP) EDNRA EDA-H52P8 Human EDNRA Full Length Protein (VLP) EDNRB EDB-H52P7 Human EDNRB Full Length Protein (VLP) EDNRB EDB-HF2G4 Fluorescent Human EDNRB Full Length Protein (VLP) GCGR GCR-H52P5 Human GCGR / Glucagon receptor Full Length Protein (VLP) GPR56 GP6-H52P7 Human GPR56 Full Length Protein (VLP) GPR87 GP7-H52P3 Human GPR87 Full Length Protein (VLP) GPRC5D GPD-H52P7 Human GPRC5D Full Length Protein (VLP) GPRC5D GPD-HF2P7 Fluorescent Human GPRC5D Full Length Protein (VLP) PAR1 PA1-H52P6 Human PAR1 Full Length Protein (VLP) PAR1 PA1-HF2G5 Fluorescent Human PAR1 Full Length Protein (VLP) PAR2 PA2-H52P7 Human PAR2 Full Length Protein (VLP) SLC7A11 & SLC3A2 SLS-H52P4 Human SLC7A11&SLC3A2 Full Length Heterodimer Protein (VLP) SMO SMO-H52P3 Human SMO Full Length Protein (VLP) SSTR2 SS2-H5216 Human SSTR2 Full Length Protein (VLP) SSTR2 SS2-HF2G3 Fluorescent Human SSTR2 Full Length Protein (VLP) VLP VLP-N5213 Virus-Like Particle (VLP) Protein Isotype Control VLP同型对照 VLP VLP-NF2P4 Fluorescent Virus-Like Particle (VLP) Protein Isotype Control 分子 货号 产品描述 应用 订购/预购 ADORA2A ADA-H52H5 Human ADORA2A Protein, His Tag (Detergent, Inhibitor added) 免疫
ELISA
SPR
BLICCR5 CC5-H52D1 Human CCR5 Protein, Flag,His Tag (Detergent) CD133 CD3-H52H4 Human CD133 Protein, His Tag (Detergent) CD20 CD0-H52D5 Human CD20 / MS4A1 Full Length Protein, Flag,His Tag (Detergent) CD20 CD0-H52H3 Human CD20 / MS4A1 Full Length Protein, His Tag (Detergent) (SPR verified) CD20 CD0-H82E5 Biotinylated Human CD20 / MS4A1 Full Length Protein, His,Avitag? (Detergent) (SPR verified) CD20 CD0-C52H8 Cynomolgus CD20 / MS4A1 Full Length Protein, His Tag (Detergent) CD20 CD0-C52H5 Canine CD20 / MS4A1 Full Length Protein, His Tag (Detergent) CD81 CD1-H85D8 Biotinylated Human CD81 Protein, His,Avitag?&Flag Tag (Detergent) Claudin-1 CL1-H55D8 Human Claudin-1 Protein, Flag,His Tag (Detergent) Claudin-18.1 CL1-H5588 Human Claudin-18.1 Protein, His,Twin-Strep Tag (Detergent) Claudin-18.2 CL2-H82E3 Biotinylated Human Claudin-18.2 Protein, His,Avitag? (Detergent) Claudin-18.2 CL2-M5585 Mouse?Claudin-18.2?Protein,?His,Twin-Strep?Tag (Detergent) Claudin-18.2 CL2-H5587 Human / Cynomolgus?Claudin-18.2?Protein,?His,Twin-Strep?Tag (Detergent) Claudin-4 CL4-H5587 Human Claudin-4 Protein, His,Twin-Strep Tag (Detergent) Claudin-6 CL6-H5289 Human Claudin-6 / CLDN6 Protein, His,Twin-Strep Tag (Detergent) Claudin-9 CL9-H5586 Human?Claudin-9 / CLDN9?Protein,?His,Twin-Strep?Tag (Detergent)?(SPR verified) CXCR3 CX3-H52D5 Human CXCR3 Protein, Flag,His Tag (Detergent) CXCR5 CX5-H52D6 Human CXCR5 Protein, Flag,His Tag (Detergent) GLP1R GLR-H52D5 Human GLP1R Protein, Flag,His Tag (Detergent) GPRC5B GPB-H52D7 Human GPRC5B Protein, Flag,His Tag (Detergent) GPRC5D GPD-H52D3 Human GPRC5D Protein, Flag,His Tag (Detergent) GPRC5D GPD-H82D6 Biotinylated Human GPRC5D Protein, His,Avitag?&Flag Tag (Detergent) GPRC5D GPD-C52D3 Cynomolgus GPRC5D Protein, Flag,His Tag (Detergent) (SPR verified) SLC1A2 SL2-H52D6 Human SLC1A2 Protein, Flag,His Tag (Detergent) STEAP1 ST1-H52H3 Human STEAP1 Full Length Protein, His Tag (Detergent) - DC-11 200x DDM CHS buffer 维持膜蛋白溶解性和活性
稀释膜蛋白或抗体
在SPR或BLI检测中制备运行缓冲液分子 货号 产品描述 应用 订购/预购 CD133 CD3-H52H1 Human CD133 Full Length Protein, His Tag (Nanodisc) 免疫
ELISA
SPR
BLI
细胞实验
CAR阳性率检测CD20 CD0-H52H1 Human CD20 / MS4A1 Full Length Protein, His Tag (Nanodisc) CD20 CD0-H82E3 Biotinylated Human CD20 / MS4A1 Full Length Protein, His,Avitag? (Nanodisc) CD20 CD0-C52H9 Canine CD20 / MS4A1 Full Length Protein, His Tag (Nanodisc) Claudin-18.2 CL2-H5586 Human?Claudin-18.2?Protein,?His,Twin-Strep?Tag (Nanodisc) Claudin-18.2 CL2-H85Q7 Biotinylated Human Claudin-18.2 Protein, Avitag?,His Tag (Nanodisc) Claudin-6 CL6-H5286 Human Claudin-6 / CLDN6 Protein,?His,Twin-Strep?Tag (Nanodisc) Claudin-6 CL6-HF2M9 Fluorescent Human Claudin-6 / CLDN6 Protein, Flag,His,GFP Tag (Nanodisc) GPRC5D GPD-H52D4 Human GPRC5D Protein, Flag,His Tag (Nanodisc) GPRC5D GPD-H82D4 Biotinylated Human GPRC5D Protein, His,Avitag?&Flag Tag (Nanodisc) GPRC5D GPD-HF2D7 FITC-Labeled Human GPRC5D Protein, Flag,His Tag (Nanodisc) GPRC5D GPD-C82D4 Cynomolgus GPRC5D Protein, Flag,His Tag (Nanodisc) MSP1D1 APO-H51H3 Human MSP1D1 Protein, His Tag (Nanodisc) 同型对照 MSP1D1 APO-H81Q5 Biotinylated Human MSP1D1 Protein, His,Avitag? (Nanodisc) 案例展示以下ELISA&SPR数据均可免费提供内部实验操作方法以供参考，点击申请Protocol即可获取。
Claudin 18.2-VLP验证数据1Claudin18.2-VLP（CL2-H52P7）
验证数据2经ELISA验证，全长Claudin18.2-VLP（CL2-H52P7）可与抗Claudin18.2抗体特异性结合，线性区间为0.2-3 ng/mL。
Immobilized Human Claudin-18.2 Full Length Protein-VLP (Cat. No. CL2-H52P7) at 5 μg/mL (100 μL/well) can bind Monoclonal Anti-Chimeric Claudin-18.2 Antibody, Human IgG1 with a linear range of 0.2-3 ng/mL (QC tested).
申请 Protocol
验证数据3经SPR验证， Claudin18.2-VLP（CL2-H52P7）可与抗Claudin18.2抗体特异性结合，亲和力常数为0.374 nM。
Human Claudin-18.2 Full Length Protein-VLP (Cat. No. CL2-H52P7) captured on CM5 Chip via Anti-Claudin-18.2 antibody can bind Anti-Claudin-18.2 antibody with an affinity constant of 0.374 nM as determined in a SPR assay (Biacore T200) (Routinely tested).
申请 Protocol咨询SPR服务验证数据4经FACS验证，用Human Claudin18.2-VLP 蛋白（Cat. No. CL2-HF218）和阴性对照蛋白（Cat. No. VLP-NF2P4）分别结合转染Claudin18.2-CAR-293细胞，
结果表明，Claudin18.2-VLP 蛋白（Cat. No. CL2-HF218）可与Claudin18.2-CAR-293细胞特异性结合。
2e5 of Anti-Claudin-18.2 CAR-293 cells were stained with 100 μL of 3 μg/mL of Fluorescent Human Claudin-18.2 Full Length Protein-VLP (Cat. No.CL2-HF218) and negative control protein respectively, FITC signals was used to evaluate the binding activity (Routinely tested).
申请 Protocol
CD20-DDM/CHS验证数据1验证数据2经ELISA验证，生物素标记全长CD20-DDM/CHS（CD0-H82E5）可与利妥昔单抗（Rituximab）特异性结合，线性区间为4-63 ng/mL。
Immobilized Rituximab at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD20 Full Length, His,Avitag (HEK293) (Cat. No. CD0-H82E5) with a linear range of 4-63 ng/mL (in presence of DDM and CHS).
申请 Protocol
验证数据3经SPR亲和力验证，生物素标记全长CD20-DDM/CHS（CD0-H82E5）可与利妥昔单抗（Rituximab）可与特异性结合，亲和力常数为1.73 nM。
Biotinylated Human CD20, His,Avitag (HEK293) (Cat. No. CD0-H82E5) captured on Biotin CAP-Series S Sensor Chip can bind Rituximab with an affinity constant of 1.73 nM as determined in a SPR assay (in presence of DDM and CHS) (Biacore T200).
申请 Protocol咨询SPR服务CD133-Nanodisc验证数据1经电泳（SDS-PAGE）验证， his标签的全长CD133-Nanodisc蛋白纯度高于90%。
Human CD133 Full Length, His Tag (Nanodisc) （(Cat. No.?CD3-H52H1)）on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.
验证数据2经ELISA验证，全长CD133-Nanodisc（CD3-H52H1）可与抗人源CD133(133-3) 单克隆抗体特异性结合，线性区间为0.2-4 ng/mL。
Immobilized Human CD133 Full Length, His Tag (Nanodisc) (Cat. No. CD3-H52H1) at 1 μg/mL (100 μL/well) can bind Monoclonal Anti-Human CD133 (133-3) Antibody, Human IgG1 with a linear range of 0.2-4 ng/mL (QC tested).
申请 Protocol
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相关参考文献Structure of CD20 in complex with the therapeutic monoclonal antibody rituximab.
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Authors: Lionel Rougé et al.
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Journal: Science
The role of CD133 in cancer: a concise review.
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Authors: Paige M. Glumac , Aaron M. LeBeau.
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Journal: Glumac and LeBeau Clin Trans Med
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Structural and Molecular Interactions of CCR5 Inhibitors with CCR5.
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Authors: Kenji Maeda, Debananda Das et al.
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Journal: Journal of Biological Chemistry
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Role of Conserved Disulfide Bridges and Aromatic Residues in Extracellular Loop 2 of Chemokine Receptor CCR8 for Chemokine and Small Molecule Binding.
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Authors: Line Barington, Pia C. Rummel, et al.
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Journal: Journal of Biological Chemistry
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Download Full Article
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