MCE 重組蛋白 凍干粉蛋白溶解方法

www.MedChemE重組蛋白R(shí)ecombinantProteins高純度

高生物活性

穩(wěn)定的一致性

GMP

級(jí)蛋白MHCVirusIgGEGFIL-6FcγRIIPD-1EGFRPD-L1IL-6RRASCARRAF-1ERK2STAT3MEK1AKT1Elk1DNAwww.MedChemEInhibitors ? ScreeningLibraries ? ProteinsMCE重組蛋白MCE重組蛋白產(chǎn)品種類豐富，涵蓋細(xì)胞因子、病毒蛋白、免疫檢查點(diǎn)蛋白、CAR-T蛋白、CD抗原、受體蛋白、生物素標(biāo)記蛋白、GMP級(jí)蛋白、酶等多種分類，可用于人、病毒、小鼠、大鼠、猴、豬、狗、牛、羊、兔、貓、非洲爪蟾等多個(gè)物種不同研究領(lǐng)域的生物學(xué)研究，包括蛋白結(jié)構(gòu)研究、免疫檢測(cè)試劑、重組蛋白藥物、診斷試劑開發(fā)、抗體藥物靶點(diǎn)、CAR-T細(xì)胞治療靶點(diǎn)、Fc受體、流感病毒蛋白和細(xì)胞因子等熱門研究領(lǐng)域。MCE因高品質(zhì)的產(chǎn)品和優(yōu)質(zhì)的售前售后服務(wù)，成為全球數(shù)百萬計(jì)的科學(xué)家和技術(shù)人員值得信賴的實(shí)驗(yàn)伙伴。產(chǎn)品種類豐富： 細(xì)胞因子和生長(zhǎng)因子、免疫檢查點(diǎn)蛋白、CAR-T相關(guān)蛋白、CD抗原、受體蛋白、酶&調(diào)節(jié)子、補(bǔ)體系統(tǒng)蛋白、病毒類蛋白、生物素標(biāo)記蛋白、熒光標(biāo)記蛋白、GMP級(jí)蛋白,泛素相關(guān)蛋白等產(chǎn)品物種多樣性：人、病毒、小鼠、大鼠、猴、豬、狗、牛、羊、兔、貓、非洲爪蟾、細(xì)菌、真菌等低內(nèi)毒素水平： LAL法驗(yàn)證，內(nèi)毒素含量低于絕大多數(shù)供應(yīng)標(biāo)準(zhǔn)高純度： SDS和HPLC檢測(cè)驗(yàn)證高生物活性： 相關(guān)體內(nèi)/體外生物學(xué)實(shí)驗(yàn)測(cè)定穩(wěn)定的一致性： 多批次檢測(cè)數(shù)據(jù)確認(rèn)規(guī)格齊全： 滿足客戶的多種預(yù)分裝規(guī)格高純度ThepurityofhumanBMP-2isgreaterthan95%asanalyzedbySDSunderreducing(R)andnon-reducing(N)condition.kDa M R 120806040302010卓越的批間穩(wěn)定性TheED50ofMCEhumanVEGF165fromthreedifferentLotsaresimilar.1.0Lot1nm)0.8Lot20.6Lot3(4900.4OD0.2Net0.01010101010VEGF165,Human(ng/mL)

高活性HumanGM-CSFstimulatescellproliferationofTF-1cellswithanED50oflessthan0.5ng/mL.2.0(490nm)1.51.0NetOD0.50.010101010101010GM-CSF,Human(ng/mL)競(jìng)品對(duì)比TheED50ofhumanVEGF165fromMCE’seachLotislowerthanofCompetitorP.1.0BrandPnm)0.8Lot20.6Lot3(4900.4OD0.2Net0.010-210-1101010VEGF165,Human(ng/mL)www.MedChemEInhibitors ? ScreeningLibraries ? Proteins凍干粉蛋白溶解方法開蓋之前需離心。凍干粉在運(yùn)輸過程中，會(huì)因?yàn)橥饬σ蛩卣掣接诠鼙诨蛘吖苌w上，在開蓋之前，先通過離心操作，將凍干粉收集于管底，避免損失和浪費(fèi)。建議10,000-12,000rpm高速離心30s；若沒有高速離心機(jī)，3,000-3,500rpm離心5min。離心之后，向凍干粉中加入提供的復(fù)溶緩沖液，用槍頭輕輕吹打混勻，重懸至濃度不低于100μg/mL。溶解過程中，不可用渦旋震蕩儀快速震蕩，否則會(huì)破壞蛋白的空間結(jié)構(gòu)，進(jìn)而影響蛋白活性。用復(fù)溶緩沖液直接溶解的重組蛋白液體可在2-8°C保存1周。對(duì)于周期較短的實(shí)驗(yàn)(不超過7天)，可以直接取該條件下保存的重組蛋白溶液加入到培養(yǎng)體系內(nèi)即可，于一周之內(nèi)用完；如果實(shí)驗(yàn)條件設(shè)置的使用濃度低于復(fù)溶之后的溶液濃度，請(qǐng)用含載體蛋白的緩沖液進(jìn)一步稀釋，如果稀釋液不含載體蛋白，容易導(dǎo)致重組蛋白粘附在管壁或者瓶壁上，使其在溶液中的濃度下降，進(jìn)一步導(dǎo)致重組蛋白的活性減弱。若要長(zhǎng)期保存，則需用含載體蛋白(0.1%BSA，5%HSA，10%FBS，或5%海藻糖)的溶液進(jìn)一步稀釋，然后分裝凍存于-20°C至-80°C。復(fù)溶后的產(chǎn)品不建議直接凍存于-20°C至-80°C。因?yàn)樗芰瞎鼙趯?duì)蛋白有很強(qiáng)的吸附作用，會(huì)導(dǎo)致重組蛋白粘附在管壁不易分離，進(jìn)而導(dǎo)致溶液中重組蛋白的實(shí)際濃度偏低，最終導(dǎo)致其活性下降。而載體蛋白可以預(yù)先封閉塑料管壁上蛋白結(jié)合位點(diǎn)，使重組蛋白不會(huì)粘附于管壁。因此長(zhǎng)期保存時(shí)，在分裝凍存前，建議用含載體蛋白的溶液進(jìn)一步稀釋(濃度不低于100μg/mL)。保存產(chǎn)品時(shí)請(qǐng)避免反復(fù)凍融反復(fù)凍融，冰晶會(huì)破壞重組蛋白的空間結(jié)構(gòu)，導(dǎo)致蛋白變性形成多聚體或使蛋白構(gòu)象發(fā)生改變，從而降低蛋白的結(jié)合能力，也加快了蛋白的降解速度。MCEMasterofBioactiveMoleculesCONTENTS細(xì)胞因子和生長(zhǎng)因子 2CytokinesandGrowthFactors免疫檢查點(diǎn)蛋白4ImmuneCheckpointProteinsCAR-T相關(guān)蛋白6CAR-TRelatedProteinsCD抗原7CDAntigensFc受體蛋白8FcReceptorProteins受體蛋白9ReceptorProteins酶&調(diào)節(jié)子10Enzymes&Regulators補(bǔ)體系統(tǒng)相關(guān)蛋白11ComplementSystemRelatedProteins泛素相關(guān)蛋白12

病毒蛋白 13ViralProteins生物素標(biāo)記蛋白 14BiotinylatedProteinsGMP級(jí)蛋白15GMP-gradeProteins無動(dòng)物成分重組蛋白 15Animal-freeRecombinantProteins重組蛋白定制服務(wù) 16RecombinantProteinExpressionService客戶驗(yàn)證 17CustomerValidation更多文獻(xiàn)引用情況 18PublicationsCitingUseofMCERecombinantProteinsUbiquitinRelatedProteinswww.MedChemEInhibitors ? ScreeningLibraries ? Proteins細(xì)胞因子和生長(zhǎng)因子細(xì)胞因子(Cytokines)是一類由多種免疫細(xì)胞(如巨噬細(xì)胞、淋巴細(xì)胞和肥大細(xì)胞等)以及某些非免疫細(xì)胞(如內(nèi)皮細(xì)胞、表皮細(xì)胞等)分泌的低分子量可溶性蛋白/多肽/糖蛋白。細(xì)胞因子包括白介素、趨化因子、干擾素等，它們?cè)谡{(diào)節(jié)細(xì)胞生長(zhǎng)、分化和激活中發(fā)揮重要作用，并參與先天和適應(yīng)性免疫反應(yīng)[1][2][3]。生長(zhǎng)因子(GrowthFactors)是一類可溶性信號(hào)分子，通常為分泌蛋白或甾體激素，能夠在發(fā)育和組織愈合過程中刺激細(xì)胞增殖、遷移、分化和多細(xì)胞形態(tài)發(fā)生[4][5]?！吧L(zhǎng)因子”和“細(xì)胞因子”常通用。某些多肽生長(zhǎng)因子為細(xì)胞因子，影響細(xì)胞生長(zhǎng)分化信號(hào)途徑的細(xì)胞因子被認(rèn)為是生長(zhǎng)因子，如GM-CSF[5][6]。MCE提供高純度、高活性細(xì)胞因子，助力您的研究！HumanIGF-I(HY-P7018)HumanIGF-I(HY-P7018)ThepurityofhumanIGF-Iisgreaterthan95%asanalyzedbySDSunderreducing(R)conditions.kDa M 120806040302010HumanEGF(HY-P7109)

TheED50ofhumanIGF-Iis<5.0ng/mlasmeasuredbyFDC-P1cells,correspondingtoaspecificactivityof>2.0×10units/mg.0.6OD4900.40.2Net0.0101010-210-11010101010IGF-I,Human(ng/mL)HumanEGF(HY-P7109)ThepurityofhumanEGFisgreaterthan95%asanalyzedbySDSunderreducing(R)conditions.kDa M R120806040302010Cat.No.ProductNameHY-P7080IL-4HY-P7223IL-6RalphaHY-P7025IFN-gammaHY-P7287SDF-1beta/CXCL12

TheED50ofhumanEGFis<0.2ng/mLasmeasuredbymurineBALB/c3T3cells,correspondingtoaspecificactivityof5.0×10units/mg.0.6OD490nm0.40.2Net0.01010101010EGF,Human(ng/mL)SpeciesSourceTagMouseMammalianCellsTag-freeHumanMammalianCellsTag-freeHumanE.coliTag-freeHumanE.coliTag-free2MCEMasterofBioactiveMoleculesCat.No.ProductNameSpeciesSourceTagHY-P7058TNF-alpha/TNFSF2HumanE.coliTag-freeHY-P7085M-CSFMouseE.coliTag-freeHY-P7118TGFbeta1HumanMammalianCellsTag-freeHY-P7086NogginMouseMammalianCellsTag-freeHY-P7007BMP-4HumanE.coliTag-freeHY-P70593FibronectinHumanE.coliTag-freeHY-P70453Wnt3aHumanMammalianCellsTag-freeHY-P7109EGFHumanE.coliTag-freeHY-P70311ActivinAHuman/Mouse/RatMammalianCellsTag-freeHY-P7319AITRL/TNFSF18MouseE.coliTag-freeHY-P75509Angiopoietin-2CanineMammalianCellsN-HisHY-P72650FGF-21CynomolgusMammalianCellsC-HisHY-P72106BMP1HumanE.coliN-HisHY-P71827Adiponectin/ADIPOQBovineP.pastorisN-HisHY-P7008BMP-7HumanE.coliTag-freeHY-P7257CCL4HumanE.coliTag-freeHY-P70450CCL5HumanE.coliTag-freeHY-P7143CCL6MouseE.coliTag-freeHY-P7772CCL9MouseE.coliTag-freeHY-P70138DLK-1HumanMammalianCellsC-HisHY-P7004FGFbasic/bFGFHumanE.coliTag-freeHY-P7170FGF-10MouseE.coliTag-freeHY-P7346FGF-8HumanE.coliTag-freeHY-P7015AG-CSFHumanMammalianCellsTag-freeHY-P7016GM-CSFHumanMammalianCellsTag-freeHY-P7017HB-EGFHumanE.coliTag-freeHY-P7018IGF-IHumanE.coliTag-freeHY-P7368IGFBP-2HumanMammalianCellsC-HisHY-P7027IL-1alphaHumanE.coliTag-freeHY-P7030AIL-10HumanMammalianCellsTag-freeHY-P7049LIFHumanE.coliTag-freeHY-P7051ANogginHumanMammalianCellsTag-freeHY-P70781SCFHumanE.coliTag-freeHY-P70467SHHHumanE.coliTag-free3www.MedChemEInhibitors ? ScreeningLibraries ? Proteins免疫檢查點(diǎn)蛋白免疫檢查點(diǎn)(ImmuneCheckpoints)分子是對(duì)免疫應(yīng)答發(fā)揮抑制或刺激作用的配體-受體對(duì)，它們?cè)诰S持免疫穩(wěn)態(tài)和免疫耐受中起著重要的調(diào)節(jié)作用。大多數(shù)被報(bào)道的免疫檢查點(diǎn)蛋白在適應(yīng)性免疫系統(tǒng)（特別是T細(xì)胞）和先天免疫系統(tǒng)的細(xì)胞上表達(dá)。一些腫瘤細(xì)胞通過結(jié)合共抑制受體分子，限制正常的抗腫瘤免疫反應(yīng)，從而幫助免疫逃逸。癌癥的免疫檢查點(diǎn)治療策略包括了靶向這些調(diào)控途徑，以恢復(fù)免疫細(xì)胞的抗腫瘤功能。目前，研究最廣泛的抑制性免疫檢查點(diǎn)包括CTLA-4和PD-1/PD-L1等[7][8]。A2aRVISTAAdenosineB7-H3PDL1/PDL2A2aRGalectin-9??CD80/CD86PD1TIM-3CD80/CD86CTLA-4OX40LCD28T-CellCD40OX40CD40LB7RP1ICOSCD70CD27HVEMBTLAAPCorLAG-3TumorCellTCRMHCl/llKIRGITRGITRL4-1BBCD226/TIGIT4-1BBLCD155/CD112OtherpotentialpathwaysIL10 CD47IDO TLRTGF-β ArginaseInhibitoryPathwaysStimulatoryPathwaysFigure1.常見的免疫檢查點(diǎn)刺激途徑和抑制途徑[9]4MCEMasterofBioactiveMoleculesCat.No.ProductNameSpeciesSourceTagHY-P70691CTLA-4HumanMammalianCellsC-GSTHY-P70632PD-L1MouseMammalianCellsC-HisHY-P7395PD-1HumanMammalianCellsC-hFcHY-P70482TIM3HumanMammalianCellsC-HisHY-P70722LAG-3HumanMammalianCellsC-HisHY-P70624TIGITHumanMammalianCellsC-HisHY-P7327CD276/B7-H3HumanMammalianCellsC-HisHY-P74464-1BBL/TNFSF9MouseMammalianCellsN-HisHY-P7144CD40L/CD154/TRAPHumanE.coliTag-freeHY-P70652CD276/B7-H3HumanMammalianCellsC-HisHY-P7678BTLA/CD272HumanMammalianCellsC-HisHY-P7685BTN3A3HumanMammalianCellsC-HisHY-P70535Galectin-9HumanMammalianCellsC-HisHY-P7366HVEMHumanSf9insectCellsC-hFcHY-P70494Nectin-1HumanMammalianCellsC-HisHY-P70807PVR/CD155HumanMammalianCellsC-HisHY-P73370PD-L2RatMammalianCellsC-hFcHY-P77879VISTAHumanMammalianCellsC-hFcHY-P75610CD137/4-1BBCanineMammalianCellsC-HisHY-P7394OX40/TNFRSF4HumanMammalianCellsC-HisHY-P77457OX40Ligand/TNFSF4CynomolgusMammalianCellsN-mFcHY-P73499CD40HumanMammalianCellsC-HisHY-P73306Nectin-3HumanMammalianCellsC-HisHY-P71248PVRIGHumanMammalianCellsC-mFcHY-P76070SIRPalphaMouseMammalianCellsC-HisHY-P73121IDOHumanE.coliTag-freeHY-P76396ICOSHumanMammalianCellsC-His-hFcHY-P77575ICOSLGCynomolgusMammalianCellsC-HisHY-P72353CD28Human/CynomolgusMammalianCellsC-Fc-AviHY-P72033LIGHTHumanMammalianCellsN-hFc-MycHY-P73076GITRHumanMammalianCellsC-HisHY-P7318GITRL/AITRLHumanE.coliTag-freeHY-P72887CD200HumanMammalianCellsC-hFcHY-P76780CD200R1CynomolgusMammalianCellsC-His5www.MedChemEInhibitors ? ScreeningLibraries ? ProteinsCAR-T相關(guān)蛋白嵌合抗原受體T(CAR-T)細(xì)胞是通過基因工程表達(dá)嵌合抗原受體的T細(xì)胞，能夠特異性地靶向抗原并殺死腫瘤細(xì)胞[10][11]。CAR-T細(xì)胞能夠通過單鏈可變區(qū)片段(scFv)結(jié)合域特異性識(shí)別靶抗原，以不依賴于主要組織相容性復(fù)合體(MHC)的方式激活，從而靶向殺傷腫瘤細(xì)胞，因此靶標(biāo)抗原的選擇對(duì)CAR-T細(xì)胞的特異性殺傷作用至關(guān)重要[12]。靶點(diǎn)蛋白結(jié)合流式細(xì)胞法是常用的CAR-T轉(zhuǎn)染陽性率檢測(cè)方案。目前MCE重組蛋白已覆蓋BCMA、EGFR、HER2、CD22、CD19、CD138、ROR1、CD123、Mesothelin等40+個(gè)CAR-T熱門靶標(biāo)。TumorCellScFvHingeRegionCostimulatoryDomainCD3ZetaFigure2.嵌合抗原受體示意圖[13]Cat.No.ProductNameSpeciesSourceTagHY-P7656BCMA/TNFRSF17MouseMammalianCellsC-FcHY-P72019Siglec-2/CD22HumanMammalianCellsC-HisHY-P70505CD19HumanMammalianCellsC-FcHY-P70731CD38HumanMammalianCellsC-HisHY-P70148MesothelinHumanMammalianCellsC-HisHY-P70189EGFRvIII`HumanMammalianCellsC-HisHY-P70125CD276/B7-H3CynomolgusMammalianCellsC-HisHY-P70155EpCAM/TROP1HumanMammalianCellsC-FcHY-P70759HER2/CD340HumanMammalianCellsC-FcHY-P70301Mucin-1HumanMammalianCellsC-FcHY-P71031Siglec-6HumanMammalianCellsC-FcHY-P70487Glypican-3/GPC3HumanMammalianCellsC-HisHY-P70296FolatereceptoralphaHumanMammalianCellsC-HisHY-P70497CD7HumanMammalianCellsC-His6MCEMasterofBioactiveMoleculesCD抗原分化簇抗原(CDAntigens)指通常用于白細(xì)胞表面分子的鑒定和研究的分化抗原群/分化抗原簇。一些CD蛋白通常充當(dāng)細(xì)胞間/細(xì)胞基質(zhì)粘附分子、細(xì)胞因子受體、離子通道或營養(yǎng)轉(zhuǎn)運(yùn)蛋白，它們?cè)诿庖呦到y(tǒng)功能中發(fā)揮多種作用。利用CD抗原作為細(xì)胞標(biāo)記物，通過熒光標(biāo)記抗體和流式細(xì)胞術(shù)結(jié)合的方法，對(duì)特定的白細(xì)胞群和淋巴細(xì)胞亞群進(jìn)行鑒定和分選[14][15]。CD45R CD45RCD28 CD28CD2 CD2CD3THTCCD3TCRTCRCD4CD8LFA1LFA1Figure3.輔助T細(xì)胞和毒性T細(xì)胞表面抗原標(biāo)志物[16]Cat.No.ProductNameSpeciesSourceTagHY-P70090CD3epsilonCynomolgusMammalianCellsC-FcHY-P72702CD8betaHumanMammalianCellsN-HisHY-P70486CD28Human/CynomolgusMammalianCellsN-HisHY-P7321B7-1/CD80HumanMammalianCellsC-hFcHY-P70029Basigin/CD147HumanMammalianCellsC-HisHY-P70507CD44HumanMammalianCellsC-HisHY-P7679BTLA/CD272HumanMammalianCellsC-FcHY-P7780Nectin-2/CD112HumanMammalianCellsC-HisHY-P7785CD127/IL-7RAHumanMammalianCellsC-Fc-HisHY-P70550CD137/4-1BBCynomolgusMammalianCellsC-HisHY-P7799CD160MouseMammalianCellsC-HisHY-P70479CD40HumanMammalianCellsC-HisHY-P7819CD207HumanMammalianCellsN-His7www.MedChemEInhibitors ? ScreeningLibraries ? ProteinsFc受體蛋白免疫球蛋白Fc受體(FcReceptors)是表達(dá)于大多數(shù)先天性和適應(yīng)性免疫細(xì)胞的膜蛋白，屬于免疫球蛋白超家族。Fc與抗體(IgA、IgD、IgE、IgG和IgM)的FabFc區(qū)域相互作用，發(fā)揮多種生物效應(yīng)，例如通過釋放細(xì)胞因子或吞噬作用調(diào)節(jié)免疫反應(yīng)。這些Fc受體包括：IgG(FcγRI/CD64、FcγRII/CD32、FcγRIII/CD16、FcRn)、IgE(FcεRI、FcεRII)、IgA(FcαRI/CD89)、IgM(FcμR)、IgD(FcδR)和Fcα/μR(IgA/IgM)等[17][18]。AntibodyD2FcReceptorEffectorCellFigure4.Fc受體與抗體包被的微生物病原體相互作用[19]

CDRVariableRegionVHVLCH1Fab-FragmentCLLightChain(korλ)CH2Fc-FragmentCH3HeavyChainsFigure5.IgG結(jié)構(gòu)圖[20]Cat.No.ProductNameSpeciesSourceTagHY-P70708FcgammaRIII/CD16MouseMammalianCellsC-HisHY-P70490FcgammaRIIIA/CD16aHumanMammalianCellsC-HisHY-P72657FCAR/CD89HumanMammalianCellsC-HisHY-P70711FcgammaRIIA/CD32aRatMammalianCellsC-HisHY-P70669CD64HumanMammalianCellsC-HisHY-P70601FCRNHumanMammalianCellsC-HisHY-P72748CD23/FcepsilonRIIHumanMammalianCellsN-HisHY-P76800FcgammaRIIB/CD32bCynomolgusMammalianCellsC-HisHY-P75204FcgammaRIIIB/CD16bHumanE.coliBiotinylatedHY-P72191FcepsilonRIA/FCER1AHumanE.coliHis-SUMOHY-P77363FCAMR/CD351MouseMammalianCellsC-HisHY-P70251IgG3FcMouseMammalianCellsTag-freeHY-P72603IgG2AFcMouseMammalianCellsTag-free8MCEMasterofBioactiveMolecules受體蛋白受體(Receptors)是位于細(xì)胞表面或胞內(nèi)的蛋白，它們與膜上或胞內(nèi)的多種信號(hào)傳導(dǎo)系統(tǒng)關(guān)聯(lián)，通過特異性地識(shí)別和結(jié)合配體調(diào)節(jié)細(xì)胞對(duì)細(xì)胞/組織微環(huán)境的反應(yīng)[21]。細(xì)胞受體種類繁多，大體分為內(nèi)部受體和細(xì)胞表面受體。細(xì)胞表面受體包括三大類：離子通道受體、G-蛋白偶聯(lián)受體(GPCRs)和酶聯(lián)受體[22]。目前，受體已經(jīng)成為一大類藥物靶點(diǎn)，例如GPCRs、細(xì)胞因子受體[23]。GLP-1-BasedDrugsGLP-1RTNF-αPalmitateLPSPKCPMAACIFN-γNLRP3cAMP/CaOx-LDLERK1/2CAMKKβPKACaspase-1pAMPK

pro-IL-1βIL-1βNF-B STAT1 NucleusCytokineProductionTNF-α,IL-6,IFN-γ,IL-1β,IL-17,IL-2

AdhesionMoleculesVCAM-1,ICAM-1,E-selectinAnti-inflammation

ChemokinesCXCL10,MCP-1InflammationInducerFigure6.基于GLP1-GLP1R分子信號(hào)的藥物抗炎作用機(jī)制[24]Cat.No.ProductNameSpeciesSourceTagHY-P70552VEGFR-2HumanMammalianCellsC-HisHY-P70714HGFRHumanMammalianCellsC-HisHY-P70352GLP1RHumanMammalianCellsC-FcHY-P70793TrkBHumanMammalianCellsC-HisHY-P7308TrkAHumanMammalianCellsTag-freeHY-P70179LIR-1/LILRB1HumanMammalianCellsC-HisHY-P7485ActivinRIB/ALK-4HumanMammalianCellsC-HisHY-P7467AGERHumanMammalianCellsC-HisHY-P71580GFRALHumanE.coliN-His-SUMOHY-P72198FSHRHumanE.coliN-HisHY-P70057TYRO3/DTKMouseMammalianCellsC-His9www.MedChemEInhibitors ? ScreeningLibraries ? Proteins酶&調(diào)節(jié)子酶類(Enzymes)是在生物系統(tǒng)內(nèi)發(fā)現(xiàn)的生物催化劑，能催化特定的生化過程。大多數(shù)酶是具有催化活性的蛋白質(zhì)，部分RNA分子(核酶)也具有催化活性。發(fā)生在生物機(jī)體內(nèi)的生物過程大多數(shù)是由酶調(diào)節(jié)的。許多遺傳性的人類疾病，如白化病和苯丙酮尿癥，都是由于缺乏特殊的酶引起的。酶催化反應(yīng)能被內(nèi)源或外源抑制劑抑制。許多毒素或活性藥物通過抑制特定的酶促反應(yīng)發(fā)揮作用[25][26][27]。1EnzymeAvailableSubstrate(sucrose)withEmptyActiveSiteActiveSite2SubstrateBindstoEnzymewithInducedFitGlucoseEnzyme(sucrase)FructoseH2O4Productsare3SubstrateisConvertedReleasedtoProductsFigure7.酶的催化作用[28]Cat.No.ProductNameSpeciesSourceTagHY-P7745CathepsinAHumanMammalianCellsC-HisHY-P7442ACE2HumanMammalianCellsC-FcHY-P70351MMP-9MouseMammalianCellsC-HisHY-P70051AminopeptidaseN/CD13MouseMammalianCellsC-HisHY-P7474ALDH1A1HumanE.coliN-HisHY-P7734CarboxypeptidaseB1/CPB1HumanMammalianCellsC-HisHY-P70005CTRB1HumanMammalianCellsC-HisHY-P70010CD73/5'-NucleotidaseHumanMammalianCellsC-HisHY-P7452ACOT13HumanMammalianCellsC-HisHY-P70221Acyl-proteinthioesterase2/LYPLA2HumanE.coliC-HisHY-P7479Aldose1-epimerase/GALMHumanE.coliC-HisHY-P72076ALOX12HumanE.coliN-HisHY-P70260Alpha-enolase/Enolase1HumanE.coliC-HisHY-P73090GSK-3betaMouseE.coliN-HisHY-P7503AngiogeninHumanE.coliTag-freeHY-P73267Kininogen-1MouseMammalianCellsC-HisHY-P7602Arginase-1HumanMammalianCellsC-His10MCEMasterofBioactiveMolecules補(bǔ)體系統(tǒng)相關(guān)蛋白補(bǔ)體系統(tǒng)(ComplementSystem)是先天免疫的主要成分之一，由可溶性血漿蛋白、膜型補(bǔ)體調(diào)節(jié)蛋白和補(bǔ)體受體組成，存在于血漿、組織、細(xì)胞表面，甚至細(xì)胞內(nèi)。補(bǔ)體系統(tǒng)主要有三條激活途徑：經(jīng)典途徑(CP)、凝集素途徑(LP)、旁路途徑(AP)。這些途徑依賴不同的起始分子激活，但最后產(chǎn)生相同的終端效應(yīng)分子：過敏性毒素(C4a/C3a/C5a)、膜攻擊復(fù)合物(MAC)和調(diào)理素(如C3b)。補(bǔ)體系統(tǒng)在防御病原體和維持宿主穩(wěn)態(tài)中起著關(guān)鍵作用[29]。ComplementSystemClassicalPathwayLectinPathwayC1qMBLAg-AbComplexesLectinBindsMannoseonPathogensC1,MASPsC4,C4C2C2CR1C3ConvertaseFcRC3C3bOpsonizationC5ConvertasePhagocytosisC5C5bC6,C7,C8C9C5b-9

AlternativePathwayPathogens,InjuredTissueC3bC3bC3aRC3aC5aRInflammationC3aRC5aC5aRMACCellLysisandActivationFigure8.三種補(bǔ)體系統(tǒng)激活途徑[30]Cat.No.ProductNameSpeciesSourceTagHY-P7863ComplementC3/C3aMouseE.coliTag-freeHY-P7864ComplementC5/C5aHumanE.coliTag-freeHY-P70031CFHR2HumanMammalianCellsC-HisHY-P70055CFHR1HumanMammalianCellsC-HisHY-P70102CFHR5HumanMammalianCellsC-HisHY-P7692HABP1/C1QBPHumanE.coliC-HisHY-P71718C1QAMouseP.pastorisN-HisHY-P71423VSIG4HumanMammalianCellsC-FcHY-P75395CD55/DAFHumanMammalianCellsC-HisHY-P7890ComplementfactorH/CFHHumanMammalianCellsC-HisHY-P74371C7/ComplementcomponentC7HumanMammalianCellsC-HisHY-P74375C2/ComplementC2HumanMammalianCellsC-HisHY-P74614ProteinS/PROS1HumanMammalianCellsC-HisHY-P75463C1s-AsubcomponentMouseMammalianCellsC-HisHY-P75464C1QBHumanSf9InsectCellsC-His11www.MedChemEInhibitors ? ScreeningLibraries ? Proteins泛素相關(guān)蛋白泛素(Ubiquitin)是真核細(xì)胞中廣泛表達(dá)的一種高度保守的小分子蛋白，由76個(gè)氨基酸組成。泛素共價(jià)結(jié)合到靶蛋白上被稱為“泛素化”，泛素化是一種重要的蛋白質(zhì)翻譯后修飾(PTM)，在控制蛋白的穩(wěn)定和功能中起重要作用，并調(diào)節(jié)了多種細(xì)胞過程，例如細(xì)胞分裂和分化、環(huán)境應(yīng)激反應(yīng)、免疫反應(yīng)、DNA修復(fù)和凋亡。著名的泛素-蛋白酶體途徑(UPP)是重要的胞質(zhì)蛋白降解途徑，能選擇性降解各種形式的受損蛋白。泛素化修飾由三類酶催化完成：1)泛素激活酶E1激活泛素蛋白；2)激活的泛素分子被轉(zhuǎn)移到泛素結(jié)合酶E2的Cys殘基上；3)泛素由E3連接酶連接到靶蛋白的Lys殘基上[31][32][33]。ATPPP1UbE1UbAMPSubstrateCysDUBAMPLysUbUbUbE1SubstrateUbCysUbE3E2E2E2E3SubstrateCysE3UbLysE1SubstrateFigure9.蛋白泛素化過程[34]Cat.No.ProductNameSpeciesSourceTagHY-P70843NEDD8HumanE.coliTag-freeHY-P71016UBE2CHumanE.coliN-HisHY-P71020UBE2L6HumanE.coliC-HisHY-P71096UCH-L1HumanE.coliC-HisHY-P71101UBBHumanE.coliTag-freeHY-P70974SUMO2HumanE.coliN-HisHY-P7617ATG3HumanE.coliTag-freeHY-P71182OTUB2HumanE.coliN-GSTHY-P71396UBAP1HumanE.coliC-HisHY-P73537XIAPHumanE.coliN-HisHY-P71643SAE1HumanE.coliN-GSTHY-P70149ISG15/UCRPHumanE.coliC-HisHY-P71395UBA5HumanE.coliN-HisHY-P71402UBE2HHumanE.coliN-GSTHY-P71407UBE2R2HumanE.coliN-His12MCEMasterofBioactiveMolecules病毒蛋白一個(gè)完整的病毒粒子由其遺傳物質(zhì)(DNA或RNA)和蛋白外殼構(gòu)成。構(gòu)成成熟的病毒粒子的蛋白，即病毒結(jié)構(gòu)蛋白，可能包括核衣殼核心蛋白(Gag蛋白)、病毒顆粒內(nèi)包裝的酶(Pol蛋白)和膜組分(Env蛋白)。病毒蛋白還包括非結(jié)構(gòu)蛋白、調(diào)節(jié)蛋白和輔助蛋白，在感染細(xì)胞時(shí)，這些蛋白在病毒復(fù)制以及組裝調(diào)節(jié)中發(fā)揮了重要作用[35][36][37]。MCE提供700+種包括了流感病毒、新型冠狀病毒(SARS-CoV-2)、人類免疫缺陷病毒(HIV)、肝炎病毒等病毒相關(guān)的重組蛋白，助力病毒研究。NucleocapsidProtein(N)MembraneGlycoprotein(M)SpikeProtein(S)EnvelopeProtein(E)RNAFigure10.SARS-CoV-2簡(jiǎn)要結(jié)構(gòu)圖[38]Cat.No.ProductNameSpeciesSourceTagHY-P74293C-likeProteinaseSARS-CoV-2E.coliN-HisHY-P7437NucleocapsidSARS-CoV-2E.coliN-HisHY-P7436S1ProteinSARS-CoV-2MammalianCellsTag-freeHY-P70127NSP1SARS-CoV-2E.coliC-HisHY-P73290Spike/S1MERS-CoVSf9insectCellsC-HisHY-P70907Envelopeglycoproteingp120HIV-1MammalianCellsC-HisHY-P74883gp140HIV-1MammalianCellsC-FcHY-P70015B18RVacciniavirusMammalianCellsC-HisHY-P71478FusionglycoproteinF0/FHRSVAE.coliN-His,B2MHY-P73232HA/HemagglutininInfluenzavirusMammalianCellsC-HisHY-P73239NA/NeuraminidaseInfluenzavirusMammalianCellsTag-freeHY-P73533MembraneproteinZikavirusMammalianCellsC-FcHY-P73738NS1ProteinDenguevirusMammalianCellsN-HisHY-P74188E/EnvelopeProteinWestNileVirusP.pastorisC-HisHY-P74354CapsidproteinHepatitisvirusE.coliC-His13www.MedChemEInhibitors ? ScreeningLibraries ? Proteins生物素標(biāo)記蛋白生物素化(Biotinylation)是將生物素以共價(jià)鍵連接到分子(如氨基酸或蛋白質(zhì))上的過程。由于生物素很小，生物素化通常快速且具有特異性，不易干擾分子固有功能。生物素與鏈霉親和素或抗生物素蛋白的結(jié)合具有極高的親和力、快速性和高特異性，因此，生物素化蛋白作為一種分子工具廣泛用于生物醫(yī)學(xué)領(lǐng)域。AviTag?是常用的融合標(biāo)簽，Avi-Tag是一種由15個(gè)氨基酸殘基組成的小標(biāo)簽，能被大腸桿菌生物素連接酶BirA有效生物素化，可添加在蛋白的N或C端，實(shí)現(xiàn)蛋白的生物素化，常用于蛋白純化或顯影等[39]。AviTagAviTagProtein—GLNDIFEAQKIEWHEProtein—GLNDIFEAQKIEWHEN3H+BirANH+OOOATPAMP+PPiSHHNHSHHNONHHHNBiotinOBiotinFigure11.Avi標(biāo)記的靶蛋白被BirA生物素化[40]Cat.No.ProductNameSpeciesSourceTagHY-P70546PCSK9HumanMammalianCellsC-His-HA-AviHY-P70721PD-L1HumanMammalianCellsC-Fc-AviHY-P70767Siglec-15HumanMammalianCellsC-Fc-AviHY-P70768CD79BHumanMammalianCellsC-His-AviHY-P70769ACE2HumanMammalianCellsC-His-AviHY-P70770TROP-2HumanMammalianCellsC-His-AviHY-P71175NTNG1HumanMammalianCellsC-Avi-HisHY-P71056TGFbeta1HumanMammalianCellsN-AviHY-P71421VEGFR-2HumanMammalianCellsC-His-AviHY-P73309Neuropilin-1HumanMammalianCellsC-His-AviHY-P73485XIAPHumanE.coliN-AviHY-P728824-1BBR/TNFRSF9HumanMammalianCellsC-hFc-AviHY-P72909CD40HumanMammalianCellsC-hFc-AviHY-P72885FcgammaRIIIA/CD16aHumanMammalianCellsC-His-AviHY-P72343BTN1A1HumanMammalianCellsC-His-AviHY-P72389LAG-3HumanMammalianCellsC-His-Avi14MCEMasterofBioactiveMoleculesGMP級(jí)蛋白GMP(生產(chǎn)質(zhì)量管理規(guī)范)重組蛋白在特定的指導(dǎo)下生產(chǎn)，以確保產(chǎn)品質(zhì)量和一致性。MCE提供的GMP級(jí)別重組蛋白是在獨(dú)立的QA監(jiān)督下生產(chǎn)，并具有質(zhì)量記錄和完全可追溯性。Cat.No.ProductNameSpeciesSourceTagHY-P70637GGMPTPOHumanMammaliancellsC-HisHY-P70757GGMPSCFHumanMammaliancellsC-HisHY-P70454GGMPIL-1alphaHumanE.coliTag-freeHY-P7044GGMPIL-6HumanE.coliTag-freeHY-P7032GGMPIL-12HumanMammalianCellsTag-freeHY-P70760GGMPIL-18HumanMammalianCellsC-HisHY-P7038GGMPIL-21HumanE.coliTag-freeHY-P78551GMPIL-2HumanE.coliTag-freeHY-P7055GGMPPDGF-BBHumanE.coliTag-free無動(dòng)物成分重組蛋白無動(dòng)物成分(Animal-free)重組蛋白，即重組蛋白生產(chǎn)過程中不使用任何動(dòng)物來源的原料和試劑，產(chǎn)品接觸的所有材料均不含任何動(dòng)物成分。在無動(dòng)物成分條件下生產(chǎn)的與使用標(biāo)準(zhǔn)實(shí)驗(yàn)室技術(shù)生產(chǎn)的蛋白具有相同的生物活性，同時(shí)避免痕量的動(dòng)物成分、動(dòng)物病原體和抗原對(duì)實(shí)驗(yàn)造成影響。Cat.No.ProductNameSpeciesSourceTagHY-P700017AFAnimal-FreeBAFFHumanE.coliC-HisHY-P700018AFAnimal-Freebeta-NGFHumanE.coliC-HisHY-P700020AFAnimal-FreeBMP-11HumanE.coliC-HisHY-P700034AFAnimal-FreeCCL2HumanE.coliN-HisHY-P700037AFAnimal-FreeCD326HumanE.coliC-HisHY-P700042AFAnimal-FreeCXCL11HumanE.coliN-HisHY-P700043AFAnimal-FreeCXCL12HumanE.coliC-HisHY-P700045AFAnimal-FreeCXCL2HumanE.coliN-HisHY-P700056AFAnimal-FreeFGF-12HumanE.coliC-HisHY-P700066AFAnimal-FreeFGF-4HumanE.coliC-HisHY-P700083AFAnimal-FreeG-CSFHumanE.coliN-His15www.MedChemEInhibitors ? ScreeningLibraries ? Proteins重組蛋白定制服務(wù)從基因設(shè)計(jì)到蛋白制備的一站式服務(wù)自主研發(fā)多種高表達(dá)載體與菌株專業(yè)的重組蛋白技術(shù)研發(fā)團(tuán)隊(duì)提供毫克至克級(jí)高純度蛋白定制服務(wù)提供蛋白在生物研究應(yīng)用中專業(yè)的技術(shù)支持

基因合成密碼子優(yōu)化交付蛋白

表達(dá)載體構(gòu)建菌株篩選蛋白表達(dá)服務(wù)內(nèi)容

蛋白純化MCE提供從基因到蛋白的一站式服務(wù)，客戶可以根據(jù)自身需求靈活性選擇服務(wù)或個(gè)性化定制服務(wù)。服務(wù)內(nèi)容包括：服務(wù)項(xiàng)目服務(wù)內(nèi)容服務(wù)周期交付內(nèi)容基因合成及亞克隆2-3周構(gòu)建報(bào)告大腸桿菌表達(dá)系統(tǒng)表達(dá)檢測(cè)1周表達(dá)檢測(cè)報(bào)告放大表達(dá)及蛋白純化1-3周純化蛋白/純化報(bào)告基因合成及亞克隆2-3周構(gòu)建報(bào)告酵母表達(dá)株電轉(zhuǎn)1-2周陽性菌株酵母細(xì)胞表達(dá)系統(tǒng)高拷貝細(xì)胞株篩選（可選）2周高拷貝菌株酵母陽性細(xì)胞株表達(dá)檢測(cè)3周表達(dá)檢測(cè)報(bào)告放大表達(dá)及蛋白純化3-5周目的蛋白/純化報(bào)告基因合成及亞克隆2-3周構(gòu)建報(bào)告哺乳動(dòng)物細(xì)胞表達(dá)系統(tǒng)瞬時(shí)轉(zhuǎn)染及表達(dá)小試2-3周蛋白表達(dá)報(bào)告放大培養(yǎng)及蛋白純化3-5周目的蛋白/純化報(bào)告基因合成及亞克隆2-3周構(gòu)建報(bào)告制備Bacmid1-2周制備報(bào)告昆蟲細(xì)胞表達(dá)系統(tǒng)桿粒及P2代病毒制備2-3周表達(dá)鑒定報(bào)告轉(zhuǎn)染及表達(dá)小試1-2周蛋白表達(dá)檢測(cè)報(bào)告放大培養(yǎng)及蛋白純化3-4周目的蛋白/純化報(bào)告16MCEMasterofBioactiveMolecules客戶驗(yàn)證誘導(dǎo)小鼠肺成纖維細(xì)胞纖維化TGF-β1(HY-P7117)刺激原代小鼠肺成纖維細(xì)胞，通過qRT-PCR和WB檢測(cè)PTPN13和NOX4表達(dá)水平，確定其與細(xì)胞纖維化密切相關(guān)[41]。ControlTGF-β1PTPN13NOX4expression10**expression8**86mRNA6mRNA44Relative2Relative200

ControlTGF-β1ControlTGF-β1CollagenI139kDaPTPN13310kDaα-SMA42kDaNOX455kDaGAPDH37kDaGAPDH37kDa構(gòu)建腎臟類器官從小鼠腎臟組織中提取細(xì)胞，利用微流體技術(shù)結(jié)合3D打印機(jī)制作類器官珠，輔以Noggin(HY-P7086)、R-spondin-1(HY-P7114)、FGF-4(HY-P7014)、FGF-basic(HY-P7066)、SB-431542(HY-10431)、Laduviglusib(HY-10182)培養(yǎng)，7天后，得到數(shù)千個(gè)大小、形狀和成分具有高度重復(fù)性的腎臟類器官[42]。OrganoidGeneratingKidneyOrganoidBeadsKidneyOrganoidsMatrigel+CellsDay1Day7QDs/Drugs7days24hrsOrganoidCulturingCulturingHighThroughputScreeningOil4°CKidneyOrganoidBeads200μm200μmCultureDish37°CIFN-γ誘導(dǎo)蛋白16(IFI16)的積累通過Westernblotting檢測(cè)IFN-γ(HY-P7025)處理人肺腺癌上皮(A549)細(xì)胞18小時(shí)后，蛋白16(IFI16)的積累，通過免疫染色檢測(cè)IFN-γ誘導(dǎo)16小時(shí)后IFI16的細(xì)胞定位[43]。HumanIFN-γ(ng/mL)0505001000IFI16—100(kDa)—70β-actin—40

NegativeControl IFN-γ(500ng/mL)DAPI/IFI1617MCEMasterofBioactiveMoleculesPublicationsCitingUseofMCEProductsNature.2023Jun;618(7964):374-382.Nature.2023Jun;618(7966):862-870.Nature.2023Jun;618(7965):590-597.Nature.2023Jun;618(7963):180-187.Nature.2023May;617(7962):818-826Science.2023Jun9;380(6649):eabo2296.Science.2022Dec2;378(6623):eabo5503.Science.2022Nov18;378(6621):eabq7361.Science.2022Oct14;378(6616):eabq0132.Science.2022Jul8;377(6602):eabg9302Cell.2023Jun22;186(13):2823-2838.e20.Cell.2023May11;186(10):2144-2159.e22.Cell.2023Mar30;186(7):1352-1368.e18.Cell.2023Mar2;186(5):1026-1038.e20.Cell.2023Feb16;186(4):850-863.e16MCEGlobalPartnerswww.MedChemEInhibitors ? ScreeningLibraries ? 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