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>产品&服务 > "膜杰作"—多次跨膜靶点蛋白开发技术平台及系列产品
Claudins家族蛋白
全长CD20蛋白
GPCRs家族蛋白

"膜杰作"—多次跨膜靶点蛋白开发技术平台及系列产品

背景
三大技术平台
产品列表
案例展示
"膜杰作"Q&A
相关参考文献
背景
多次跨膜靶点蛋白（ACROBiosystems均可提供）
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
VLP技术平台
VLP技术平台
查看更多详情
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
膜蛋白-去垢剂技术平台
膜蛋白-去垢剂技术平台
查看更多详情
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
Nanodisc技术平台
Nanodisc技术平台
查看更多详情
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 订购/预购
ADORA2A ADA-H52P4 Human ADORA2A Full Length Protein (VLP)

订购

CCR4 CC4-H52P3 Human CCR4 Full Length Protein (VLP)

订购

CCR5 CC5-H52P5 Fluorescent Human CCR5 Full Length Protein (VLP)

订购

CCR5 CC5-H52P3 Human CCR5 Full Length Protein (VLP)

订购

CCR6 CC6-H52P3 Human CCR6 Full Length Protein (VLP)

订购

CCR7 CC7-H52P3 Human CCR7 Full Length Protein (VLP)

订购

CCR8 CC8-H52P4 Human CCR8 Full Length Protein (VLP)

订购

CD133 CD3-H52P3 Human CD133 Full Length Protein (VLP)

订购

CD151 CD1-H52P9 Human CD151 Full Length Protein (VLP)

订购

CD20 CDP-HF2P8 Fluorescent Human CD20 Full Length Protein (VLP)

订购

CD20 CDP-H52P6 Human CD20 Full Length Protein (VLP)

订购

CD20 CD0-C52P3 Canine CD20 Full Length Protein (VLP)

订购

CD37 CD7-H52P4 Human CD37 Full Length Protein (VLP)

订购

CD63 CD3-H52P6 Human CD63 Full Length Protein (VLP)

订购

CD81 CD1-H52P5 Human CD81 Full Length Protein (VLP)

订购

CD9 CD9-H52P3 Human CD9 Full Length Protein (VLP)

订购

CGRPR & RAMP1 CG1-H52P6 Human CGRPR&RAMP1 Full Length Heterodimer Protein (VLP)

订购

Claudin-1 CL1-H52P6 Human Claudin-1 Full Length Protein (VLP)

订购

Claudin-18.1 CL1-H52P3 Human Claudin-18.1 Full Length Protein (VLP)

订购

Claudin-18.2 CL2-H52P7 Human Claudin-18.2 Full Length Protein (VLP)

订购

Claudin-18.2 CL2-HF218 Fluorescent Human Claudin-18.2 Full Length Protein (VLP)

订购

Claudin-3 CL3-H52P3 Human Claudin-3 Full Length Protein (VLP)

订购

Claudin-4 CL4-H52P5 Human Claudin-4 Full Length Protein (VLP)

订购

Claudin-6 CL6-HF2G8 Fluorescent Human Claudin-6 / CLDN6 Full Length Protein (VLP)

订购

Claudin-6 CL6-H52P7 Human Claudin-6 / CLDN6 Full Length Protein (VLP)

订购

Claudin-9 CL9-H52P5 Human Claudin-9 / CLDN9 Full Length Protein (VLP)

订购

CNR1 CN1-H52P6 Human CNR1 Full Length Protein (VLP)

订购

CXCR3 CX3-H52P4 Human CXCR3 Full Length Protein (VLP)

订购

CXCR3 CX3-M52P7 Mouse CXCR3 Full Length Protein (VLP)

订购

CXCR4 CX4-H5219 Human CXCR4 / CD184 Full Length Protein (VLP)

订购

CXCR5 CX5-H52P4 Human CXCR5 Full Length Protein (VLP)

订购

CXCR6 CX6-H52P3 Human CXCR6 / CD186 Full Length Protein (VLP)

订购

EDNRA EDA-H52P8 Human EDNRA Full Length Protein (VLP)

订购

EDNRB EDB-H52P7 Human EDNRB Full Length Protein (VLP)

订购

EDNRB EDB-HF2G4 Fluorescent Human EDNRB Full Length Protein (VLP)

订购

GCGR GCR-H52P5 Human GCGR / Glucagon receptor Full Length Protein (VLP)

订购

GPR56 GP6-H52P7 Human GPR56 Full Length Protein (VLP)

订购

GPR87 GP7-H52P3 Human GPR87 Full Length Protein (VLP)

订购

GPRC5D GPD-H52P7 Human GPRC5D Full Length Protein (VLP)

订购

GPRC5D GPD-M52P3 Mouse GPRC5D Full Length Protein (VLP)

订购

LIV-1 LV1-H52P3 Human LIV-1 Full Length Protein (VLP)

订购

LPAR1 LP1-H52P3 Human LPAR1 Full Length Protein (VLP)

订购

PAR1 PA1-HF2G5 Fluorescent Human PAR1 Full Length Protein (VLP)

订购

PAR1 PA1-H52P6 Human PAR1 Full Length Protein (VLP)

订购

PAR2 PA2-H52P7 Human PAR2 Full Length Protein (VLP)

订购

SLC7A11 & SLC3A2 SLS-H52P4 Human SLC7A11&SLC3A2 Full Length Heterodimer Protein (VLP)

订购

SMO SMO-H52P3 Human SMO Full Length Protein (VLP)

订购

SSTR2 SS2-H5216 Human SSTR2 Full Length Protein (VLP)

订购

SSTR2 SS2-HF2G3 Fluorescent Human SSTR2 Full Length Protein (VLP)

订购

VLP VLP-N5213 Virus-Like Particle (VLP) Protein Isotype Control

订购

VLP VLP-NF2P4 Fluorescent Virus-Like Particle (VLP) Protein Isotype Control

订购

分子 货号 产品描述 订购/预购
5-HT2AR 5HR-H55D3 Human 5-HT2AR Protein (S162K, M164W, S372N), Flag,His Tag (Detergent)

订购

ADORA2A ADA-H52D3 Human ADORA2A Protein, Flag,His Tag (Detergent)

订购

C5AR1 C51-H52D3 Human C5AR1 Protein, Flag,His Tag (Detergent)

订购

CCR4 CC4-H52D3 Human CCR4 Full Length Protein, Flag,His Tag (Detergent)

订购

CCR5 CC5-H52D1 Human CCR5 Protein, Flag,His Tag (Detergent)

订购

CCR5 CC5-M52D9 Mouse CCR5 Protein, Flag,His Tag (Detergent)

订购

CCR7 CC7-H82D4 Biotinylated Human CCR7 Full Length Protein, His,Flag Tag&Avitag™ (Detergent)

订购

CCR7 CC7-H52D3 Human CCR7 Full Length Protein, His,Flag Tag (Detergent)

订购

CCR8 CC8-H52D6 Human CCR8 Protein, Flag,His Tag&Twin-Strep Tag (Detergent)

订购

CCR8 CC8-H82D3 Biotinylated Human CCR8 Protein, His,Flag Tag&Avitag™ (Detergent)

订购

CCR9 CC9-H82D3 Human CCR9 Protein, His,Avitag™&Flag Tag (Detergent)

订购

CCR9 CC9-H52D3 Human CCR9 Full Length Protein, Flag,His Tag (Detergent)

订购

CD133 CD3-H52H4 Human CD133 Protein, His Tag (Detergent)

订购

CD133 CD3-H82E5 Biotinylated Human CD133 Full Length Protein, His,Avitag™ (Detergent)

订购

CD133 CD3-C82E3 Biotinylated Cynomolgus CD133 Full Length Protein, His,Avitag™ (Detergent)

订购

CD151 CD1-H52D3 Human CD151 Protein, Flag,His Tag (Detergent)

订购

CD20 CD0-H82E5 Biotinylated Human CD20 / MS4A1 Full Length Protein, His,Avitag™ (Detergent) (SPR verified)

订购

CD20 CD0-H52H3 Human CD20 / MS4A1 Full Length Protein, His Tag (Detergent) (SPR verified)

订购

CD20 CD0-H52D5 Human CD20 / MS4A1 Full Length Protein, Flag,His Tag (Detergent)

订购

CD20 CD0-C52H8 Cynomolgus CD20 / MS4A1 Full Length Protein, His Tag (Detergent)

订购

CD20 CD0-C52H5 Canine CD20 / MS4A1 Full Length Protein, His Tag (Detergent)

订购

CD20 CD0-C52Hb Cat CD20 / MS4A1 Full Length Protein, His Tag (Detergent)

订购

CD63 CD3-H55D3 Human CD63 Full Length Protein, Flag,His Tag (Detergent)

订购

CD81 CD1-H85D8 Biotinylated Human CD81 Protein, His,Avitag™&Flag Tag (Detergent)

订购

Claudin-1 CL1-H55D8 Human Claudin-1 Protein, Flag,His Tag (Detergent)

订购

Claudin-18.1 CL1-H5588 Human Claudin-18.1 Protein, His,Twin-Strep Tag (Detergent)

预购

Claudin-18.2 CL2-H85D3 Biotinylated Human Claudin-18.2 Protein, His,Avitag™&Flag Tag (Detergent)

订购

Claudin-18.2 CL2-M5585 Mouse Claudin-18.2 Protein, His,Twin-Strep Tag (Detergent)

订购

Claudin-18.2 CL2-H5587 Human / Cynomolgus Claudin-18.2 Protein, His,Twin-Strep Tag (Detergent)

订购

Claudin-4 CL4-H5587 Human Claudin-4 Protein, His,Twin-Strep Tag (Detergent)

订购

Claudin-6 CL6-H85D3 Biotinylated Human Claudin-6 Protein, His,Avitag™&Flag Tag (Detergent)

订购

Claudin-6 CL6-H5289 Human Claudin-6 / CLDN6 Protein, His,Twin-Strep Tag (Detergent)

订购

Claudin-9 CL9-H5586 Human Claudin-9 / CLDN9 Protein, His,Twin-Strep Tag (Detergent)

订购

CNR1 CN1-H55D3 Human CNR1 Protein, Flag,His Tag (Detergent)

订购

CXCR3 CX3-H52D5 Human CXCR3 Protein, Flag,His Tag (Detergent)

订购

CXCR4 CX4-H52D3 Human CXCR4 Full Length Protein, Flag,His Tag (Detergent)

订购

CXCR5 CX5-H52D6 Human CXCR5 Protein, Flag,His Tag (Detergent)

订购

GCGR GCR-H52D3 Human GCGR Protein, Flag,His Tag (Detergent)

订购

GIPR GIR-H52D3 Human GIPR Protein, His,Flag Tag (Detergent)

订购

GLP1R GLR-H52D4 Human GLP1R Full Length Protein (L260F), Flag,His Tag (Detergent)

订购

GPR75 GP5-H52D3 Human GPR75 Full Length Protein, Flag,His Tag (Detergent)

订购

GPRC5B GPB-H52D7 Human GPRC5B Protein, Flag,His Tag (Detergent)

订购

GPRC5D GPD-H82D5 Biotinylated Human GPRC5D Full Length Protein, Flag,His,Avitag™ (Detergent)

订购

GPRC5D GPD-H52D5 Human GPRC5D Full Length Protein, Flag,His Tag (Detergent)

订购

GPRC5D GPD-C52D3 Cynomolgus GPRC5D Protein, Flag,His Tag (Detergent) (SPR verified)

订购

SLC1A2 SL2-H52D6 Human SLC1A2 Protein, Flag,His Tag (Detergent)

订购

SLC2A1 SL1-H55H3 Human SLC2A1 Full Length Protein, His Tag (Detergent)

订购

SLC7A11 & SLC3A2 SLS-H52D3 Human SLC7A11&SLC3A2 Full Length Protein, Flag,His Tag (Detergent)

订购

STEAP1 ST1-H82D3 Biotinylated Human STEAP1 Full Length Protein, Avitag™,Flag&His Tag(Detergent)

订购

STEAP1 ST1-H5283 Human STEAP1 Full Length Protein, Twin-Strep,His Tag (Detergent)

订购

TM4SF5 TM5-H52D3 Human TM4SF5 Protein, Flag,His Tag (Detergent)

订购

- DC-11 200x DDM CHS buffer

订购

- DN-12 50x Digitonin buffer

订购

- LG-13 200x LMNG CHS buffer

订购

分子 货号 产品描述 订购/预购
CCR8 CC8-H52D7 Human CCR8 Protein, Flag,His Tag&Twin-Strep Tag (Nanodisc)

订购

CD133 CD3-H52H1 Human CD133 Full Length Protein, His Tag (Nanodisc)

订购

CD20 CD0-H52H1 Human CD20 / MS4A1 Full Length Protein, His Tag (Nanodisc)

订购

CD20 CD0-H82E3 Biotinylated Human CD20 / MS4A1 Full Length Protein, His,Avitag™ (Nanodisc)

订购

CD20 CD0-HF2H5 FITC-Labeled Human CD20 / MS4A1 Full Length Protein, His Tag (Nanodisc, SPR verified)

订购

CD20 CD0-CF2H3 FITC-Labeled Cynomolgus CD20 / MS4A1 Full Length Protein, His Tag (Nanodisc)

订购

CD20 CD0-C52H9 Canine CD20 / MS4A1 Full Length Protein, His Tag (Nanodisc)

订购

Claudin-18.2 CL2-H85D4 Biotinylated Human Claudin-18.2 Protein, His,Avitag™&Flag Tag (Nanodisc)

订购

Claudin-18.2 CL2-H5586 Human / Cynomolgus Claudin-18.2 Protein, His,Twin-Strep Tag (Nanodisc)

订购

Claudin-6 CL6-H5286 Human Claudin-6 / CLDN6 Protein, His,Twin-Strep Tag (Nanodisc)

订购

Claudin-6 CL6-HF2M9 Fluorescent Human Claudin-6 / CLDN6 Protein, Flag,His,GFP Tag (Nanodisc)

订购

GPRC5D GPD-HF2D6 FITC-Labeled Human GPRC5D Full Length Protein, Flag,His Tag (Nanodisc)

订购

GPRC5D GPD-H82D7 Biotinylated Human GPRC5D Full Length Protein, Flag,His,Avitag™ (Nanodisc)

订购

GPRC5D GPD-H52D6 Human GPRC5D Full Length Protein, Flag,His Tag (Nanodisc)

订购

GPRC5D GPD-C82D4 Cynomolgus GPRC5D Protein, Flag,His Tag (Nanodisc)

订购

MSP1D1 APO-H81Q5 Biotinylated Human MSP1D1 Protein, His,Avitag™ (Nanodisc)

订购

MSP1D1 APO-H51H3 Human MSP1D1 Protein, His Tag (Nanodisc)

订购

我们还可提供相应稳定细胞株及细胞株开发服务，助力抗体药物开发及临床申报。欢迎咨询~
案例展示
Claudin 18.2-VLP Claudin 18.2-VLP
CD20-DDM/CHS CD20-DDM/CHS
CD133-Nanodisc CD133-Nanodisc

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1
Claudin18.2-VLP（CL2-H52P7）
Claudin 18.2-VLP
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。
Claudin 18.2-VLP

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。
Claudin 18.2-VLP

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。
Claudin 18.2-VLP

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 (QC tested).

申请 Protocol

CD20-DDM/CHS
验证数据1
经ELISA验证，全长CD20-DDM/CHS（CD0-H52H3）可与利妥昔单抗（Rituximab）特异性结合，线性区间为0.4-3 ng/mL。
CD20-DDM/CHS

Immobilized Rituximab at 2 μg/mL (100 μL/well) can bind Human CD20 Full Length Protein, His Tag (Cat. No. CD0-H52H3) with a linear range of 0.4-3 ng/mL (in presence of DDM and CHS) (QC tested).

申请 Protocol

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。
CD20-DDM/CHS

Immobilized Rituximab at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD20 Full Length, His,Avitag (Cat. No. CD0-H82E5) with a linear range of 4-63 ng/mL (in presence of DDM and CHS) (QC tested).

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。
CD20-DDM/CHS

Biotinylated Human CD20 Full Length, His,Avitag (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) (QC tested).

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%。
  
CD133-Nanodisc

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。
CD133-Nanodisc

Immobilized Human CD133 Full Length Protein, His Tag (Cat. No. CD3-H52H1) at 1 μg/mL (100 μL/well) can bind Monoclonal Anti-Human CD133 Antibody, Human IgG1 with a linear range of 0.2-4 ng/mL (QC tested).

申请 Protocol

Q1：如何解读“膜杰作”？

"膜杰作"一方面是指平台，ACRO突破了技术难题，搭建了三大技术平台，为多次跨膜蛋白的成功研发提供了平台保障。另一方面，"膜杰作"是指ACRO依托于平台研发出的多次跨膜蛋白产品，具有全长、天然构象、稳定、高活性、覆盖多应用场景等杰出优势。 "杰"，也是ACRO的"技术自信"，以杰出的研发能力，服务于更好的生物医药。

Q2：想购买Claudin18.2做免疫和抗体筛选，怎么挑选？

我们制备的Claudin18.2有VLP平台下的版本，也有DDM/CHS平台下的Claudin18.2版本。两款都是可以的。不过免疫的话也可以优先选择VLP版本的，因为本身VLP可以提高免疫原性。筛选的话两款都是可以，如果想要得到准确的亲和力数据，推荐DDM/CHS版本（即去垢剂版本）。

Q3：抗体与抗原结合在胞外，全长的多次跨膜蛋白意义大吗？

多次跨膜蛋白在细胞膜上多次跨越，在胞外形成多个胞外区，比如Claudin18.2、CD20、CD133都有两个ECL（extracellular loops），每个ECL都有特定功能甚至会相互影响。全长可以保证蛋白构象天然完整，而且ECL展现完全，帮助筛选靶向多次跨膜蛋白抗原的理想抗体。给大家看下面两张图，全长的意义就显而易见了，结构完整，功能上也有保证。这么看，全长也不是我们ACRO一直在强调全长，而是客户需要全长的多次跨膜蛋白，满足客户需要的产品是ACRO的初衷。我们也比较过，单独的胞外ECL区蛋白活性确实比全长蛋白差。

多次跨膜蛋白就是要全长
Q4：VLP和Nanodisc平台下的多次跨膜蛋白怎么去除非特异抗体？

非特异抗体的影响不容忽视，使用VLP和Nanodisc平台下的膜蛋白的确可能产生非特异抗体，我们这两个平台均有对应的用于反筛去除非特异抗体的空白对照：VLP-膜蛋白有同型对照产品空白VLP；Nanodisc有两款MSP1D1空白产品，其中一款MSP1D1作为tag free 膜蛋白的同型对照，如果使用生物素化的Nanodisc-膜蛋白，另一款MSP1D1可以作为同型对照。

VLP/Nanodisc 同型对照产品
Q5：相比其他非Nanodisc产品，Nanodisc有什么好处吗？

Nanodisc-膜蛋白与去垢剂-膜蛋白相比，的确有不同。首先，原理上，Nanodisc膜蛋白是将膜蛋白除去去垢剂之后组装在类细胞膜的Nanodisc上。其次，应用上，我们都知道，去垢剂会溶膜从而对细胞产生损伤，Nanodisc版本的膜蛋白因不含去垢剂，可以进一步扩展到用于细胞试验、CAR阳性率检测，应用范围是更广的。另外，我们ACRO使用的Nanodisc技术已经获得专利持有方的授权，在研发过程中是可以放心使用的。

Q6：如何确认VLP-Claudin18.2表达的是该目标蛋白，纯度如何评估？产品上的电镜图片，怎么确认颗粒上有该蛋白?

Claudin18.2-VLP 我们会通过anti-Claudin18.2特异性的抗体进行检测验证。纯度我们通过电泳/HPLC/DLS电镜进行评估。常规负染电镜因分辨率低是没办法看到VLP上是否有Claudin18.2的。

Q7：VLP相关蛋白能适合冻干保存吗？VLP的用于免疫有什么注意事项？比如能否乳化，对佐剂类型有没有要求？

包膜VLP产品我们目前都是液体-70°C保存，干冰发货；非包膜VLP我们是冻干保存的。VLP免疫主要是需要注意佐剂量的选择，因为VLP本身是可以增强免疫原性的，跟常规蛋白不同。

Q8： CD24，PD-1等，也有VLP产品？和Claudin18.2是一样的吗？包膜VLP和非包膜有什么区别?

CD24，PD-1本身属于可溶蛋白，不是多次跨膜蛋白，所以使用的是非包膜的VLP；而多次跨膜蛋白因为有疏水跨膜区，所以需要使用包膜VLP；两者的区别就是VLP的表面是否有磷脂双分子膜。

Q9：VLP和Nanodisc产品稳定性如何。冷冻长时间保存可以吗？

VLP和Nanodisc本身的稳定性是很好的，是可以在-70°C长期保存的。

Q10：定制的全长多次跨膜蛋白，多久能到货？

定制的话，VLP版本的膜蛋白预计需要6-8周；DDM/CHS去垢剂版本的膜蛋白需要进行多次跨膜靶点蛋白表达纯化条件摸索和去垢剂筛选，所以开发周期更长一些，需要8周以上；Nanodisc版本的膜蛋白是DDM/CHS的周期上再加4周左右。

ACRO膜蛋白技术分享交流群聚焦多次跨膜靶点蛋白研发、应用等相关主题的讨论，热点文章分享和解读，欢迎大家热烈参与讨论、积极分享。
识别下方二维码，添加ACRO微信小助手，加入膜蛋白技术分享群吧~
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相关参考文献

Structure of CD20 in complex with the therapeutic monoclonal antibody rituximab.

  • Authors: Lionel Rougé et al.

  • Journal: Science

The role of CD133 in cancer: a concise review.

  • Authors: Paige M. Glumac , Aaron M. LeBeau.

  • Journal: Glumac and LeBeau Clin Trans Med

Structural and Molecular Interactions of CCR5 Inhibitors with CCR5.

  • Authors: Kenji Maeda, Debananda Das et al.

  • Journal: Journal of Biological Chemistry

Role of Conserved Disulfide Bridges and Aromatic Residues in Extracellular Loop 2 of Chemokine Receptor CCR8 for Chemokine and Small Molecule Binding.

  • Authors: Line Barington, Pia C. Rummel, et al.

  • Journal: Journal of Biological Chemistry

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