DiMPro?功能性膜蛋白開發(fā)平臺(tái)

跨膜蛋白作為細(xì)胞膜的重要組成部分, 在物質(zhì)運(yùn)輸、信號(hào)轉(zhuǎn)導(dǎo)和細(xì)胞間識(shí)別等多種細(xì)胞功能中發(fā)揮著重要作用。其功能異常往往會(huì)導(dǎo)致疾病的發(fā)生，這使它們成為理想的藥物作用靶點(diǎn)。目前以跨膜蛋白為藥物靶點(diǎn)占現(xiàn)階段己知藥物靶點(diǎn)的60%以上。而針對(duì)抗體藥靶點(diǎn)，膜蛋白幾乎占90%以上。盡管有著重大的意義，針對(duì)跨膜蛋白的藥物開發(fā)仍然具有很大的挑戰(zhàn)性，主要是因?yàn)榭缒さ鞍椎氖杷Y(jié)構(gòu)，使得其在體外很難以可溶性蛋白形式保持其天然構(gòu)象，同時(shí)全長(zhǎng)多跨膜蛋白通常表達(dá)水平較低，這些都是跨膜蛋白表達(dá)的難點(diǎn)所在。如何獲得最為天然構(gòu)象和具有功能活性的膜蛋白是開發(fā)這一類靶點(diǎn)抗體藥物的核心所在。

締碼生物科技有限公司為了促進(jìn)藥物研發(fā)，自主研發(fā)出多種針對(duì)膜蛋白的制備方案。利用HEK293細(xì)胞表達(dá)系統(tǒng)表達(dá)出更接近天然蛋白構(gòu)象的重組膜蛋白。對(duì)于單穿膜蛋白，主要采取胞外結(jié)構(gòu)域(ECD)融合蛋白表達(dá)系統(tǒng)。而針對(duì)全長(zhǎng)多穿膜蛋白，如GPCR和Claudin系列蛋白，則采用締碼七大全長(zhǎng)多跨膜蛋白表達(dá)平臺(tái)，包括純化膜蛋白平臺(tái)和非純化膜蛋白平臺(tái)。純化膜蛋白平臺(tái)包括：Synthetic Nanodisc、MSP Nanodisc、PeptiNanodisc和去垢劑平臺(tái)。非純化膜蛋白平臺(tái)包括：MNP(納米膜顆粒)、VLP和外泌體平臺(tái)。

DiMProm Membrane Protein Technology Platforms

平臺(tái)介紹

胞外結(jié)構(gòu)域(ECD)融合蛋白表達(dá)平臺(tái)

胞外域蛋白表達(dá)平臺(tái)
  • 適用于單穿膜蛋白靶點(diǎn)
  • 1000+個(gè)現(xiàn)貨的ECD重組蛋白
  • HEK293哺乳動(dòng)物表達(dá)系統(tǒng)
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全長(zhǎng)多跨膜蛋白表達(dá)平臺(tái)

純化膜蛋白平臺(tái)

純化膜蛋白平臺(tái)
純化膜蛋白平臺(tái)區(qū)別

非純化膜蛋白平臺(tái)

非純化膜蛋白平臺(tái)
非純化膜蛋白平臺(tái)

平臺(tái)流程

膜蛋白平臺(tái)流程

平臺(tái)優(yōu)勢(shì)

  1. 哺乳動(dòng)物表達(dá)系統(tǒng)可表達(dá)出最接近天然蛋白構(gòu)象及翻譯后修飾的重組膜蛋白
  2. 無血清培養(yǎng)體系：最大限度地減少宿主細(xì)胞污染
  3. 功能活性驗(yàn)證
  4. 可實(shí)現(xiàn)蛋白定制和放大生產(chǎn)

關(guān)鍵應(yīng)用

  1. 用于治療性抗體藥物開發(fā)的活性免疫原
  2. 配體受體結(jié)合實(shí)驗(yàn)
  3. 藥物的臨床前體外功能性實(shí)驗(yàn)
  4. 蛋白質(zhì)功能測(cè)試
  5. 基于細(xì)胞學(xué)的功能檢測(cè)試驗(yàn)

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