DiMPro™功能性膜蛋白开发平台

跨膜蛋白作为细胞膜的重要组成部分, 在物质运输、信号转导和细胞间识别等多种细胞功能中发挥着重要作用。其功能异常往往会导致疾病的发生，这使它们成为理想的药物作用靶点。目前以跨膜蛋白为药物靶点占现阶段己知药物靶点的60%以上。而针对抗体药靶点，膜蛋白几乎占90%以上。尽管有着重大的意义，针对跨膜蛋白的药物开发仍然具有很大的挑战性，主要是因为跨膜蛋白的疏水结构，使得其在体外很难以可溶性蛋白形式保持其天然构象，同时全长多跨膜蛋白通常表达水平较低，这些都是跨膜蛋白表达的难点所在。如何获得最为天然构象和具有功能活性的膜蛋白是开发这一类靶点抗体药物的核心所在。

缔码生物科技有限公司为了促进药物研发，自主研发出多种针对膜蛋白的制备方案。利用HEK293细胞表达系统表达出更接近天然蛋白构象的重组膜蛋白。对于单穿膜蛋白，主要采取胞外结构域(ECD)融合蛋白表达系统。而针对全长多穿膜蛋白，如GPCR和Claudin系列蛋白，则采用缔码七大全长多跨膜蛋白表达平台，包括纯化膜蛋白平台和非纯化膜蛋白平台。纯化膜蛋白平台包括：Synthetic Nanodisc、MSP Nanodisc、PeptiNanodisc和去垢剂平台。非纯化膜蛋白平台包括：MNP(纳米膜颗粒)、VLP和外泌体平台。

DiMProm Membrane Protein Technology Platforms

平台介绍

胞外结构域(ECD)融合蛋白表达平台

胞外域蛋白表达平台
  • 适用于单穿膜蛋白靶点
  • 1000+个现货的ECD重组蛋白
  • HEK293哺乳动物表达系统
点击查看服务详情

全长多跨膜蛋白表达平台

纯化膜蛋白平台

纯化膜蛋白平台
纯化膜蛋白平台区别

非纯化膜蛋白平台

非纯化膜蛋白平台
非纯化膜蛋白平台

平台流程

膜蛋白平台流程

平台优势

  1. 哺乳动物表达系统可表达出最接近天然蛋白构象及翻译后修饰的重组膜蛋白
  2. 无血清培养体系：最大限度地减少宿主细胞污染
  3. 功能活性验证
  4. 可实现蛋白定制和放大生产

关键应用

  1. 用于治疗性抗体药物开发的活性免疫原
  2. 配体受体结合实验
  3. 药物的临床前体外功能性实验
  4. 蛋白质功能测试
  5. 基于细胞学的功能检测试验

案例展示

Human BCMA Protein, mFc Tag (PME100035)

Human BCMA, mFc Tag on SDS-PAGE under reducing condition

Human BAFF protein (100 μl/per well) (PME100043) binds to Human BCMA protein (PME100035) (EC50 0.03-15.625 ng/ml).

uploads-PME100035 BCMA mFc ELISA Fig31 2

Human BCMA protein (2 μg/ml) (PME100035) binds to Anti-BCMA Neutralizing antibody (BME100028) (EC50 0.64-80.0 ng/ml).

FC analysis with 1μg/ml Human BCMA Protein on HEK293 cells transfected with human BAFF (Blue) or HEK293 transfected with irrelevant protein (Red).

Human GPR75-Strep full length protein-synthetic nanodisc (FLP120031)

Human GPR75-Strep-Nanodisc on SDS-PAGE

WB analysis of GPR75-Strep-Nanodisc with anti-GPR75 mAb (DMC100368)

Human GPR75-Strep-Nanodisc (0.2μg/per well) binds to anti-GPR75 mAb (DMC100368) (EC50 15.14ng/ml).

Loaded Human GPR75 mAb (DMC100368) on Pro-A Biosensor, binds human GPR75-Strep-synthetic nanodisc ( an affinity constant 5.02nM) .

Human CLDN18.2-Strep full length protein-PeptiNanodisc (FLP420014)

SDS-PAGE Analysis the Purity of CLDN18.2-PeptiNanodisc

CLDN18.2-Peptidisc (0.2μg/per well) binds anti-CLDN18.2 mAb (DME100179) (EC50 60.34ng/ml). 

Human CLDN6 full length protein -MNP (FLP100004)

Human CLDN6 membrane nanoparticles (0.5μg/per well) binds to anti-CLDN6 mAb (BME100082) (EC50 34.36ng/ml).

FACS analysis of CLDN6 MNPs

A. CLDN6 MNPs were stained only with secondary Ab. B. Control MNPs (2μg/mL) were stained with anti-CLDN6 antibody (BME100082), followed by secondary Ab. C. CLDN6 MNPs (2μg/mL) were stained with anti-GPRC5D antibody, followed by secondary Ab. D. CLDN6 MNPs (2μg/mL) were stained with anti-CLDN6 antibody (BME100082), followed by  secondary Ab.

Human CLDN18.2 full length protein-VLP (FLP100006)

Human CLDN18.2 VLP (0.5ug/per well) binds to Anti-CLDN18.2 mAb (Zolbetuximab biosililar) (EC50 15.37ng/ml).

FACS analysis of CLDN18.2 VLP

A. CLDN18.2 VLP were stained only with PE secondary Ab.
B. Control VLP (1μg/mL) were stained with anti-CLDN18.2 antibody (Zolbetuximab biosimilar), followed by PE secondary Ab.
C. CLDN18.2 VLP (1μg/mL) were stained with anti-BCMA antibody, followed by PE secondary Ab.
D. CLDN18.2 VLP (1μg/mL) were stained with anti-CLDN18.2 antibody（Zolbetuximab biosimilar) , followed by PE secondary Ab.

Human CD24 full length protein-exo (FLP100002)

Human CD24 exosome (0.5μg/per well) binds to Anti-CD24 mAb (EC50 69.61ng/ml).

Nanoparticle Tracking Analysis of CD24 exosomes

TEM image of CD24 exosomes