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      技術(shù)文章您現(xiàn)在的位置：首頁 > 技術(shù)文章 > InVivoKines™體內(nèi)研究專用蛋白：超高活性和穩(wěn)定性
       
      
      
        
      InVivoKines™體內(nèi)研究專用蛋白：超高活性和穩(wěn)定性
      
        
      更新時間：2025-01-24      點擊次數(shù)：413次
      
        
      細(xì)胞因子是一類小蛋白，能在細(xì)胞間傳遞信息，具有免疫調(diào)節(jié)和效應(yīng)功能，這些作用使得它們在各種免疫相關(guān)疾病的臨床前和臨床研究中極-具吸引力。它們在免疫中發(fā)揮關(guān)鍵作用，而基于工程化細(xì)胞因子的療法代表了免疫治療的新發(fā)展。然而細(xì)胞因子的廣泛應(yīng)用受到血液半衰期短、多效性和脫靶效應(yīng)的限制。對細(xì)胞因子生物學(xué)的深入了解以及創(chuàng)新的細(xì)胞因子工程技術(shù)可以應(yīng)對這些限制，但他們通常又無法用于基礎(chǔ)研究。
       
      InVivoKines™ 是 AdipoGen Life Sciences利用Knobs-into-Holes (KIH) 技術(shù)研發(fā)的新一代重組融合蛋白，用于免疫治療、臨床前和轉(zhuǎn)化體內(nèi)研究。如需訂購Adipogen公司產(chǎn)品，請聯(lián)系國內(nèi)授權(quán)代理欣博盛生物。
      InVivoKines™ 優(yōu)勢與特點：
      • 具有天然構(gòu)象——在HEK293或CHO細(xì)胞中生產(chǎn)
      穩(wěn)定性.png
      • 在無動物條件下生產(chǎn)
      • 通過ELISA及基于細(xì)胞的檢測測試的高生物活性
      • 通過SEC驗證的純度和均一性
      • 低內(nèi)毒素水平
      • 批次間一致性

      天然細(xì)胞因子與Fc融合(KIH技術(shù))
      InVivoKines™ 是使用 Knobs-into-Holes (KIH) 技術(shù)研發(fā) 的Fc融合蛋白
      • Fc-KIH技術(shù)允許Fc異源二聚體化，創(chuàng)造出具有兩個不同臂的結(jié)構(gòu)，以設(shè)計天然存在的活性單體、異源二聚體或同源二聚體蛋白
      • Fc-KIH 域增強了細(xì)胞因子的血漿半衰期，從而在保持活性的同時顯著改善藥代動力學(xué)(PK)
      • Fc-KIH 域與 Fcγ受體(FcγRs)和補體 C1q結(jié)合，可能導(dǎo)致抗體依賴性細(xì)胞介導(dǎo)的細(xì)胞毒性作用(ADCC)、抗體依賴性細(xì)胞介導(dǎo)的吞噬作用(ADCP)和補體依賴的細(xì)胞毒性作用(CDC)
      • Fc-KIH LALA-PG突變抑制了與FcγRs和C1q的結(jié)合，而與FcRn結(jié)合和Fc穩(wěn)定性不受影響
      • Fc-KIH 在內(nèi)毒素含量低的哺乳動物細(xì)胞中生產(chǎn)
      • Fc-KIH 蛋白是為體內(nèi)研開的，但也同樣適用于體外實驗！

      AdipoGen Life Sciences利用KIH技術(shù)開發(fā)單體、異源二聚體或同源二聚體Fc融合蛋白.
      穩(wěn)定性-1.png
      沉默的Fc-KIH域(LALA-PG 突變)
      AdipoGen Life Sciences利用hIgG1-P329G LALA突變來設(shè)計其沉默的Fc-KIH域，這些突變完-全消除了與FcγR和C1q的相互作用，包含了有限數(shù)量的突變，并且不影響FcRn的相互作用和 Fc 的穩(wěn)定性。LALA-PG突變顯示未檢測到與Fc受體或 C1q 的結(jié)合，在基于功能細(xì)胞的檢測中無活性，并且不引發(fā)炎癥性細(xì)胞因子反應(yīng)。
      穩(wěn)定性-2.png
      LITERATURE:
      •  (1) Knobs-into-Holes (KIH) Technology: J.B. Ridgway, et al.; Protein Eng. 9, 617 (1996) 
      • (2) Efficient Generation of Bispecific Murine Antibodies for Pre-Clinical Investigations in Syngeneic Rodent Models: A.F. Labrijn, et al.; Sci. Rep. 7, 2476 (2017) 
      • (3) Novel human IgG1 and IgG4 Fc-engineered antibodies with completely abolished immune effector functions:T. Schlothauer, et al.; Prot. Eng. Design Sel. 29, 457 (2016)
       
      單體IL-2蛋白
      IL-2 Superkine已被證明能誘導(dǎo) T 細(xì)胞(CD4/CD8)增殖(非 T 調(diào)節(jié)細(xì)胞)，導(dǎo)致細(xì)胞毒性CD8 T細(xì)胞和NK細(xì)胞的顯著擴增，因此，與人IL-2 WT相比，體內(nèi)抗腫瘤反應(yīng)得到改善。IL-2 Superkine(單體):Fc-KIH比IL-2 Superkine:Fc(二聚體)更高效，等同于或優(yōu)于金標(biāo)準(zhǔn) IL-2 Aldesleukin。
      IL-2 Superkine (monomeric): Fc-KIH (human)(rec.)
      穩(wěn)定性-3.png
      AG-40B-022210μg|3x10μg|100μg
      NEWIL-2 Superkine H9T (monomeric): Fc-KIH(human) (rec.)
      AG-40B-022310μg|3x10μg|100μg
      IL-2 (human) (monomeric): Fc-KIH (human) (rec.)
      AG-40B-022410μg|3x10μg|100μg
      IL-2 (mouse) (monomeric): Fc-KIH (human) (rec.)
      AG-40B-022510μg|3x10μg|100μg
      FIGURE: IL-2 Superkine (monomeric):Fc-KIH (human) (rec.) (Prod. No. AG-40B-0222)activates the IL-2 receptor better than the dimeric IL-2 Superkine ((Prod. No. AG40B-0111) using the IL-2 Bioass ay (Promega JA2201).

      NEW   IL-27 (mouse):Fc-KIH (human) (rec.)      AG-40B-0236
      IL-27由兩個亞基IL-27p28和EBI3組成。它促進(jìn)NK和T細(xì)胞增殖以及 IFN-γ 的產(chǎn)生。IL-27對多種病毒具有強大的抗病毒活性，并通過產(chǎn)生抗血管生成趨化因子具有抗腫瘤活性。

      IL-27 (mouse):Fc-KIH (human) (rec.)是一種異源二聚體Fc-KIH結(jié)構(gòu)體，與小鼠和人的IL-27R復(fù)合物結(jié)合，激活小鼠和人細(xì)胞中Stat3磷酸化。與傳統(tǒng)的二聚體IL-277 (mouse):Fc相比，其活性更強。
      穩(wěn)定性-4.png
      FIGURE: IL-27 (mouse):Fc-KIH (human) (rec.) (Prod. N o.AG-40B-0236) (Heterodimer)is more active than an IL-27 (mouse):Fc from a competitor (Dimer of Heterodimer)in Stat 3 phosphorylation in HepG2 cells (shown as quantification figure of the westernblot)
       
      NEW Fc (LALA-PG)-KIH (human):GDF15 (mouse) (rec.)      AG-40B-0245
      GDF15的作用是多方面的，包括對食欲調(diào)節(jié)、代謝、妊娠、細(xì)胞存活、免疫反應(yīng)和炎癥的作用。

      AdipoGen Life Sciences的Fc (LALAPG)-KIH(human):GDF15(mouse) (rec.)是一種在體內(nèi)研究中具有長效高活性的獨-特蛋白質(zhì)，與其他形成無活性大聚集體的經(jīng)典GDF15-Fc蛋白不同，是具有生物活性的同源二聚體。
      穩(wěn)定性-5.png
      FIGURE: A binding assay shows that Fc (LALA-PG)-KIH (human):GDF15 (mouse) (rec.) (Prod. No. AG-40B-0245) binds with high affinity to its receptor GFRAL (mouse). The Fc (LALA-PG)-KIH(human):GDF15 (mouse) (rec.), which migrates as a dimer by SEC, binds to its receptor mouse GFRAL with an EC50 ~15 ng/ml, while the GDF15 (mouse):Fc (human) (rec.), that migrates asan aggregate by SEC (>1000 kDa), binds similar to the control protein with low affinity.

      InVivoKines™?產(chǎn)品目錄
      產(chǎn)品名稱貨號結(jié)構(gòu)Fc-KIH 默認(rèn)
      IL-2 (human) (monomeric): Fc-KIH (human) (rec.)AG-40B-0224MonomerNo
      IL-2 (mouse) (monomeric): Fc-KIH (human) (rec.)AG-40B-0225MonomerNo
      IL-2 Superkine (monomeric): Fc-KIH (human) (rec.)AG-40B-0222MonomerNo
      IL-2 Superkine H9T (monomeric): Fc-KIH (human) (rec.)AG-40B-0223MonomerNo
      IL-2 (human) (Switch-2) (monomeric): Fc-KIH (human) (rec.)AG-40B-0234MonomerNo
      IL-7 (human) (monomeric): Fc-KIH (human) (rec.)AG-40B-0238MonomerNo
      IL-7 (human) (monomeric): Fc (LALA-PG)-KIH (human) (rec.) NEWAG-40B-0247MonomerLALA-PG
      IL-12 (mouse): Fc-KIH (human) (rec.)AG-40B-0240HeterodimerNo
      IL-21 (mouse) (monomeric): Fc (LALA-PG)-KIH (human) (rec.) NEWAG-40B-0250MonomerLALA-PG
      IL-23 (mouse): Fc-KIH (human) (rec.)AG-40B-0235HeterodimerNo
      IL-23 (mouse): Fc (LALA-PG)-KIH (human) (rec.) NEWAG-40B-0248HeterodimerLALA-PG
      IL-27 (mouse): Fc-KIH (human) (rec.)AG-40B-0236HeterodimerNo
      IL-27 (mouse): Fc (LALA-PG)-KIH (human) (rec.) NEWAG-40B-0249HeterodimerLALA-PG
      IL-33 (oxidation resistant) (human) (monomeric): Fc-KIH (human) (rec.)AG-40B-0233MonomerNo
      IL-37 (human) (monomeric): Fc-KIH (human) (rec.)AG-40B-0221MonomerNo
      IL-38 (aa 1-152) (human) (monomeric): Fc-KIH (human) (rec.)AG-40B-0241MonomerNo
      IL-38 (aa 20-152) (human) (monomeric): Fc-KIH (human) (rec.)AG-40B-0226MonomerNo
      IL-38 (aa 3-152) (mouse) (monomeric): Fc-KIH (human) (rec.)AG-40B-0227MonomerNo
      Fc (LALA-PG)-KIH (human): GDF15 (mouse) (rec.) NEWAG-40B-0245HomodimerLALA-PG
      Fc (LALA-PG)-KIH (human): Irisin (monomeric) (rec.) NEWAG-40B-0246MonomerLALA-PG
      Fc-KIH (human) IgG1 Control (rec.)AG-35B-0015ControlNo
      Fc (LALA-PG)-KIH (human) IgG1 Control (rec.)AG-35B-0018ControlLALA-PG

      InVivoKines™ —可選結(jié)構(gòu)
      穩(wěn)定性-6.png

      17313129617344465.png
      更多詳情，請聯(lián)系A(chǔ)dipoGen中國授權(quán)代理商——欣博盛生物