多次跨膜蛋白
多次跨膜蛋白是细胞生长、信号转导及细胞间通讯的重要功能载体,广泛参与机体的各项生理活动,同时也是药物开发的重要靶点。然而,多次跨膜蛋白一直以来存在可溶重组形式的制备难题。恺佧生物拥有完善的VLP和Nanodisc技术平台,实现了多次跨膜蛋白的可溶表达,不仅保持了多次跨膜蛋白正确的结构和构象,且具备优异的生物学活性,可助力多次跨膜蛋白的相关药物研发。
VLP展示多次跨膜蛋白
VLP即Virus Like Particle,是由一种或多种病毒衣壳蛋白自动组装形成的微小纳米颗粒,具有很好的包封分子的能力。VLP展示多次跨膜蛋白是将多次跨膜蛋白展示在VLP表面,不仅维持了多次跨膜蛋白的正确构象,同时具备较强的免疫原性,在抗体发现中具有极大优势。

VLP展示多次跨膜蛋白机理

恺佧生物成功开发出一系列VLP展示的多次跨膜蛋白产品,靶点涵盖Claudin 18.2、GPRC5D、Claudin 6、CD20等多种靶点,突破了此类蛋白制备困难、可溶性低的技术壁垒,且具有更强的免疫原性,产品均经过严格的生物学活性验证,可满足客户不同场景需求。同时,为了实现SPR等更多场景的开发应用,恺佧生物特别开发出定点生物素化的VLP超级蛋白产品,具有高灵敏度、高特异性及高稳定性特点,适合多种分析方法。
产品优势:
全长序列表达,具备天然结构和构象
强免疫原性
可定点引入生物素化
全面的活性验证
产品应用:
动物免疫
抗体淘选
体内药效药代研究
ELISA、SPR等各种分析检测
数据示例
Fig. 1 Immobilized Human Claudin 18.2 VLP at 5 μg/ml (100 μl/Well). Dose response curve for Anti-Claudin 18.2 Antibody, hFc Tag with the EC50 of 9.8 ng/ml determined by ELISA.
Fig. 2 Biotinylated Human Claudin 18.2 VLP captured on CM5 Chip via Streptavidin can bind Anti-Claudin 18.2 Antibody with an affinity constant of 1.28 nM as determined in SPR assay.
Fig. 3 Immobilized Human Claudin 6 VLP at 5 μg/ml (100 μl/Well). Dose response curve for Anti-Claudin 6 Antibody, mFc Tag with the EC50 of 2.5 ng/ml determined by ELISA.
Fig. 4 Biotinylated Human Claudin 6 VLP captured on CM5 Chip via Streptavidin can bind Anti-Claudin 6 Antibody with an affinity constant of 0.65 nM as determined in SPR assay.
Nanodisc展示多次跨膜蛋白
多次跨膜蛋白的结构和功能与其所处的膜环境息息相关,多次跨膜蛋白研究的发展方向之一是开发能够提供更好的细胞膜模拟效果的纯化方法,Nanodisc展示技术应运而生。

Nanodisc展示多次跨膜蛋白示意图

Nanodisc也称纳米圆盘,为一种由磷脂和膜支架分子组成的类膜结构,是用于研究膜蛋白结构和功能的重要工具。它通过模拟天然磷脂双分子层膜环境,为膜蛋白提供了一个稳定的平台,利用Nanodisc展示多次跨膜蛋白不仅可以更好地维持蛋白的天然构象,还可解决多次跨膜蛋白的可溶性制备问题,广泛应用于药物开发的各种分析检测过程。
产品特点
正确的结构和构象
非去垢剂提取,性能更可靠
定量更精准
产品应用
抗体淘选
抗体筛选
检测方法开发(ELISA/SPR/BLI等)
数据示例
恺佧生物开发的Nanodisc展示全长GPRC5D蛋白具有优异的生物学活性和较高的检测灵敏度。
Fig. 5 Immobilized Human GPRC5D Nanodisc, His Tag at 2 μg/ml (100 μl/well) on the plate. Dose response curve for Anti-GPRC5D Antibody, hFc Tag with the EC50 of 4.9 ng/ml determined by
Fig. 6 Human GPRC5D Nanodisc, His Tag captured on CM5 Chip via anti-his antibody can bind Anti-GPRC5D Antibody with an affinity constant of 1.47 nM as determined in SPR assay.
为更大程度助益抗体药物(如双抗药物)开发,恺佧生物特别提供生物素化的Nanodisc展示多次跨膜蛋白,在测评中表现出优异的结合活性,可灵活应用于ELISA方法开发及药代动力学分析等不同研究阶段。
Fig. 7 Immobilized Human CD3E&CD3D, His Tag at 2 μg/ml (100 μl/well) on the plate, add serial dilutions of Anti-Human CD3×GPRC5D Bispecific Antibody, hFc Tag and then add Biotinylated Human GPRC5D Nanodisc, His Tag at 5 μg/ml. Detection was performed using HRP-conjugated streptavidin with the EC50 of 0.28 μg/ml determined by ELISA.
Fig. 8 Loaded Biotinylated Human GPRC5D Nanodisc, His Tag on SA-Biosensor, can bind Anti-GPRC5D Antibody with an affinity constant of 1.16 nM as determined in BLI assay.
多次跨膜蛋白在抗体药物PK分析中的应用
多次跨膜蛋白的靶向药物开发是目前的热门研究方向,在其抗体相关药物的PK分析中,可运用多次跨膜蛋白对抗体进行亲和捕获,以分析抗体的浓度、活性等不同参数,比如在ADC药物的LC-MS分析中,能分析总抗体和偶联抗体的浓度及DAR等,进而反映药物的药效或毒性。

多次跨膜蛋白的PK分析应用举例:ADC药物的LC-MS分析原理

Fig. 9 Immobilized Human CD3E&CD3D at 5 μg/ml (100 μl/well) on the plate, add serial dilutions of Anti-Human CD3XGPRC5D bispecific antibody in 60% human serum and then add various dilution ratio of biotinylated human GPRC5D VLP. Detection was performed using HRP-conjugated streptavidin with the EC50 of 0.28 μg/ml determined by ELISA. The backcalculation bias is less than 20%, and the sensitivity of the method can reach 6.6 ng/ml.
Fig. 10 lmmobilized Human Claudin 6 VLP at 2 μg/ml (100 μl/well) on the plate. Add serial dilutions of Anti-Claudin 6 Antibody in 60% human serum. Detection was performed using HRP-conjugated goat anti-mouse antibody with the EC50 of 4.5 ng/ml determined by ELISA. The back calculation bias is less than 25%, and the sensitivity of the method can reach 0.7 ng/ml.
蛋白名称 货号 产品名称 标签 表达系统
A2AR A2R-HM0P108 Human A2AR VLP N/A HEK293 立即询价
A2R-HM1N1 Human A2AR Nanodisc C-His HEK293 立即询价
APLNR APL-HM0P112 Human APLNR VLP N/A HEK293 立即询价
CB1 CNR-HM001 Human Cannabinoid receptor 1 VLP N/A HEK293 立即询价
CCR2b CCR-HM02B Human CCR2b VLP N/A HEK293 立即询价
CCR-HM02BB Biotinylated Human CCR2b VLP N/A HEK293 立即询价
CCR4 CR4-HM1N122 Human CCR4 Nanodisc C-His HEK293 立即询价
CCR6 CCR-HM16NB Biotinylated Human CCR6 Nanodisc C-His HEK293 立即询价
CCR7 CCR-HM107 Human CCR7 Nanodisc C-His HEK293 立即询价
CCR-HM17NB Biotinylated Human CCR7 Nanodisc C-His HEK293 立即询价
CCR8 CCR-HM08V Human CCR8 VLP N/A HEK293 立即询价
CR8-HM1N29 Human CCR8 Nanodisc C-His HEK293 立即询价
CD133 CD33-HM0P110 Human CD133 VLP N/A HEK293 立即询价
CD20 CD2-HM123 Human CD20/MS4A1 VLP N/A HEK293 立即询价
CD2-HM123B Biotinylated Human CD20/MS4A1 VLP N/A HEK293 立即询价
CD2-HM12NB Biotinylated Human CD20/MS4A1 Nanodisc C-His HEK293 立即询价
Claudin 18.2 CLD-HM0P37 Human Claudin 18.2 VLP N/A HEK293 立即询价
CLD-HE1822 Human Claudin 18.2 VLP N/A HEK293 立即询价
CLD-HE1822B Biotinylated Human Claudin 18.2 VLP N/A HEK293 立即询价
CLD-HM10NB Biotinylated Human Claudin 18.2 Nanodisc C-His HEK293 立即询价
Claudin 3 CLD-HM13NB Biotinylated Human Claudin 3 Nanodisc C-His HEK293 立即询价
Claudin 4 CLD-HM104 Human Claudin 4 VLP N/A HEK293 立即询价
CLD4-HM1N161 Human Claudin 4 Nanodisc C-His HEK293 立即询价
CLD-HM14NB Biotinylated Human Claudin 4 Nanodisc C-His HEK293 立即询价
Claudin 6 CLD-HM006 Human Claudin 6 VLP N/A HEK293 立即询价
CLD-HM006B Biotinylated Human Claudin 6 VLP N/A HEK293 立即询价
CLD-CM006 Cynomolgus Claudin 6 VLP N/A HEK293 立即询价
CLD-MM006 Mouse Claudin 6 VLP N/A HEK293 立即询价
CLD-HM16NB Biotinylated Human Claudin 6 Nanodisc C-His HEK293 立即询价
Claudin 9 CLD-HM009 Human Claudin 9 VLP N/A HEK293 立即询价
CX3CR1 CXR-HM00V Human CX3CR1 VLP N/A HEK293 立即询价
CXR-HM10NB Biotinylated Human CX3CR1 Nanodisc C-His HEK293 立即询价
CXCR4 CXR-HM1RNB Biotinylated Human CXCR4 Nanodisc C-His HEK293 立即询价
CXCR5 CXC-HM1R5 Human CXCR5 VLP N/A HEK293 立即询价
CXR5-HM1N72 Human CXCR5 Nanodisc C-His HEK293 立即询价
CXC-HM1RNB Biotinylated Human CXCR5 Nanodisc C-His HEK293 立即询价
SLC7A11 XCT-HM10NB Biotinylated Human Cystine/glutamate transporter (SLC7A11) Nanodisc C-His HEK293 立即询价
EDNRA EDA-HM0P111 Human EDNRA VLP N/A HEK293 立即询价
GCGR GCR-HM00V Human GCGR/Glucagon receptor VLP N/A HEK293 立即询价
GCR-HM10NB Biotinylated Human GCGR/Glucagon receptor Nanodisc C-His HEK293 立即询价
GIPR GIP-HM14RNB Biotinylated Human GIPR Nanodisc C-His HEK293 立即询价
GLP-1R GLP-HM1N123 Human GLP-1R Nanodisc C-His HEK293 立即询价
GLP-HM10NB Biotinylated Human GLP-1R Nanodisc C-His HEK293 立即询价
GPRC5D GPR-HM05P Human GPRC5D VLP N/A HEK293 立即询价
GPR-HM05PB Biotinylated Human GPRC5D VLP N/A HEK293 立即询价
GPR-CM05P Cynomolgus GPRC5D VLP N/A HEK293 立即询价
GPR-MM05P Mouse GPRC5D VLP N/A HEK293 立即询价
GPR-HM15P Human GPRC5D Nanodisc C-His HEK293 立即询价
GPR-HM15NB Biotinylated Human GPRC5D Nanodisc C-His HEK293 立即询价
LGR-4 LGR-HM10N Human LGR-4 Nanodisc C-His HEK293 立即询价
LGR-5 LR5-HM1N157 Human LGR-5 Nanodisc C-His HEK293 立即询价
LPAR1 LPA-HM0P111 Human LPAR1/LPA receptor 1 VLP N/A HEK293 立即询价
MRGPRX2 MR2-HM00V Human MRGPRX2 VLP N/A HEK293 立即询价
MR2-HM1N118 Human MRGPRX2 Nanodisc C-His HEK293 立即询价
MR2-HM10XN Biotinylated Human MRGPRX2 Nanodisc C-His HEK293 立即询价
Polycystin-2 PC2-HM10N Human PC2/Polycystin-2 Nanodisc C-His HEK293 立即询价
PAR-1 PAR-HM01R Human Proteinase-activated receptor 1/PAR-1 VLP N/A HEK293 立即询价
SSTR2 STR-HM002 Human SSTR2 VLP N/A HEK293 立即询价
STEAP1 STP-HM01V Human STEAP1 VLP N/A HEK293 立即询价
STP-HM00VB Biotinylated Human STEAP1 VLP N/A HEK293 立即询价
TM4SF1 TSF-HM002 Human TM4SF1 VLP N/A HEK293 立即询价
TSF-HM00N Human TM4SF1 Nanodisc N-His HEK293 立即询价
TSHR THR-HM00V Human TSHR VLP N/A HEK293 立即询价
Control VLP-HM00C VLP Control N/A HEK293 立即询价
GPR-HM05CB Biotinylated VLP Control N/A HEK293 立即询价
CON-HM1NB Biotinylated Nanodisc Control C-His HEK293 立即询价