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IL-2及IL-2受体蛋白

IL-2对免疫反应具有多效作用
背景
白细胞介素(ILs)是一类细胞因子,最初被发现由白细胞表达,目前已发现至少超过38种。白细胞介素2(Interleukin-2,IL-2)是最早发现的细胞因子之一,又称为T细胞生长因子(T cell growth factor, TCGF),是一种重要的免疫调节因子。
IL-2是最早的一类免疫治疗药物,但其必须以很高的剂量才能产生效果,且毒性很强,会引发致命的血管泄漏综合征和其它副作用。之后,有研究发现,IL-2其实具有双重活性。高剂量的IL-2能促进T细胞增殖,驱动抗肿瘤活性,但也会激活调节性T细胞(Tregs);低剂量IL-2仅激活调节性T细胞,抑制免疫细胞的免疫反应,可以治疗自身免疫性疾病。
IL-2的受体interleukin-2 receptor(IL-2R)包含α、β和γ三个亚单位。三个亚单位可以组成不同亲和力的受体,其中,单独的IL-2R α为低亲和力受体,单独存在不参与信号传导,IL-2R β和IL-2R γ组成中等亲和力的受体,下游可以通过JAK传导信号,在IL-2R α参与的情况下,亲和力增强为高亲和力受体。IL-2与其受体特异性结合后,启动Jak-STAT、MAPK及P13K等多种途径,抑制凋亡,诱导细胞增殖、分化,产生多种生物学效应。基于以上结构生物学的研究,不同研发公司进行了不同IL-2改造和设计的方法,包括PEG化修饰、融合Fc、IL-2突变体设计、双特异性抗体设计、联合免疫检查点抗体用药等策略。因此,结构稳定的高亲和力IL-2受体复合物,对于体外研究IL-2与IL-2受体的相互作用、免疫动物制备抗体或抗体的体外筛选等具有重要意义。
天然IL-2对免疫反应具有多效作用

图1 天然IL-2对免疫反应具有多效作用[1]

IL-2与低亲和力、中等亲和力和高亲和力IL-2受体结合的示意图

图2 IL-2与低亲和力、中等亲和力和高亲和力IL-2受体结合的示意图[2]

然而,由于IL-2受体复合物结构的特殊性,α位于IL-2的一侧,而β和γ形成“Y”形,位于IL-2的另一侧[3],在没有IL-2存在时,α、β和γ无法直接形成高亲和力的IL-2受体复合物。这也限制了针对IL-2Rαβγ异源三聚体的抗体制备等应用。
ACROBiosystems开发了IL-2及其受体蛋白:IL-2IL-2RαIL-2RβIL-2RγIL-2Rβγ异源二聚体蛋白,以及采用自有专利技术研发IL-2Rαβγ异源三聚体蛋白,可用于研究IL-2与IL-2受体的相互作用、免疫动物制备抗体或抗体的体外筛选等,助力IL-2相关药物研发。此外我们还可以提供 GMP级别IL-2产品,可用于T/NK/iPSC等多种细胞的规模化培养。
产品列表
  • IL-2
  • IL-2Rα
  • IL-2Rβ
  • IL-2Rγ
  • IL-2Rβγ
  • IL-2Rαβγ
分子 货号 种属 标签 产品描述
IL-2 CRS-A003 Human Protein-coupled magnetic beads resDetect™ Human Interleukin-2 (IL-2) ELISA Kit (Residue Testing)
CRS-B008 Human Protein-coupled magnetic beads ClinMax™ Human IL-2 ELISA Kit
GMP-L02H14 Human Tag Free GMP Human IL-2 Protein
IL2-H5215 Human Tag Free Human IL-2 Protein (HEK293), premium grade
IL2-H5269 Human Fc Tag Human IL-2 Protein, Fc Tag (MALS verified)
IL2-H52H8 Human His Tag Human IL-2 Protein, His Tag
IL2-H82E4 Human His Tag & Avi Tag Biotinylated Human IL-2 Protein, His,
IL2-H82F3 Human Fc Tag & Avi Tag Biotinylated Human IL-2 Protein, Fc,
IL2-M52H3 Mouse His Tag Mouse IL-2 Protein, His Tag
IL2-C5249 Cynomolgus His Tag Cynomolgus IL-2 Protein, His Tag
分子 货号 种属 标签 产品描述
IL-2 R alpha ILA-H5251 Human Fc Tag Human IL-2 R alpha / CD25 Protein, Fc Tag (MALS verified)
ILA-H52H9 Human His Tag Human IL-2 R alpha / CD25 Protein, His Tag
ILA-H82E6 Human His Tag & Avi Tag Biotinylated Human IL-2 R alpha / CD25 Protein, His,, premium grade
ILA-H82F9 Human Fc Tag & Avi Tag Biotinylated Human IL-2 R alpha / CD25 Protein, Fc, (MALS verified)
ILA-M52H9 Mouse His Tag Mouse IL-2 R alpha / CD25 Protein, His Tag
ILA-C52H8 Cynomolgus His Tag Cynomolgus IL-2 R alpha / CD25 Protein, His Tag (MALS verified)
ILA-C52H4 Canine His Tag Canine IL-2 R alpha / CD25 Protein, His Tag
分子 货号 种属 标签 产品描述
IL-2 R beta CD2-H5221 Human His Tag Human IL-2 R beta / CD122 Protein, His Tag (SPR verified)
ILB-H5253 Human Fc Tag Human IL-2 R beta / CD122 Protein, Fc Tag (MALS & SPR verified)
ILB-H82E3 Human His Tag & Avi Tag Biotinylated Human IL-2 R beta / CD122 Protein, His,
ILA-M52H5 Mouse His Tag Mouse IL-2 R beta Protein, His Tag
ILB-M5253 Mouse Fc Tag Mouse IL-2 R beta / CD122 Protein, Fc Tag
ILB-C52H9 Cynomolgus His Tag Cynomolgus IL-2 R beta / CD122 Protein, His Tag (MALS verified)
ILA-C52H5 Canine His Tag Canine IL-2 R beta Protein, His Tag
分子 货号 种属 标签 产品描述
IL-2 R gamma ILA-H52H5 Human His Tag Human IL-2 R gamma / CD132 Protein, His Tag (MALS verified)
ILG-H5256 Human Fc Tag Human IL-2 R gamma / CD132 Protein, Fc Tag
ILG-H85E8 Human His Tag & Avi Tag Biotinylated Human IL-2 R gamma / CD132 Protein, His,
ILA-M52H3 Mouse His Tag Mouse IL-2 R gamma / CD132 Protein, His Tag (MALS verified)
ILA-M82E3 Mouse His Tag & Avi Tag Biotinylated Mouse IL-2 R gamma Protein, His, (MALS verified)
ILG-C52H9 Cynomolgus His Tag Cynomolgus IL-2 R gamma / CD132 Protein, His Tag (MALS verified)
分子 货号 种属 标签 产品描述
IL-2 R beta & IL-2 R gamma CHEK-ATF136 Human Protein-coupled magnetic beads Human IL-2 R beta/IL-2 R gamma (Luc) HEK293 Reporter Cell
ILG-H5254 Human Fc Tag Human IL-2 R beta&IL-2 R gamma Heterodimer Protein, Fc Tag&Fc Tag (MALS verified)
ILG-H5283 Human His Tag & Strep Tag Human IL-2 R beta&IL-2 R gamma Heterodimer Protein, His Tag&Twin-Strep Tag (MALS verified)
ILG-H82F3 Human Fc Tag & Avi Tag Biotinylated Human IL-2 R beta&IL-2 R gamma Heterodimer Protein, Fc,&Fc, (MALS verified)
ILG-M5254 Mouse Fc Tag Mouse IL-2 R beta&IL-2 R gamma Heterodimer Protein, Fc Tag&Fc Tag (MALS verified)
ILG-M5283 Mouse His Tag & Strep Tag Mouse IL-2 R beta&IL-2 R gamma Heterodimer Protein, His Tag&Twin-Strep Tag (MALS verified)
ILG-C5283 Cynomolgus His Tag & Strep Tag Cynomolgus IL-2 R beta&IL-2 R gamma Heterodimer Protein, His Tag&Twin-Strep Tag (MALS verified)
ILG-C82F4 Cynomolgus Fc Tag & Avi Tag Biotinylated Cynomolgus IL-2 R beta&IL-2 R gamma Heterodimer Protein, Fc,&Fc, (MALS verified)
ILG-R5283 Rat His Tag & Strep Tag Rat IL-2 R beta&IL-2 R gamma Heterodimer Protein, His Tag&Twin-Strep Tag (MALS verified)
分子 货号 种属 标签 产品描述
IL-2 R beta & IL-2 R alpha & IL-2 R gamma ILG-H5257 Human Fc Tag Human IL-2 R beta&IL-2 R alpha&IL-2 R gamma Protein, Fc Tag&Fc Tag (MALS verified)
ILG-H52W9 Human His Tag & Strep Tag Human IL-2 R beta&IL-2 R alpha&IL-2 R gamma Protein, His Tag&Twin Strep Tag (MALS verified)
ILG-H82F4 Human Fc Tag & Avi Tag Biotinylated Human IL-2 R beta&IL-2 R alpha&IL-2 R gamma Protein, Fc,&Fc, (MALS verified)
ILG-H82W9 Human Protein-coupled magnetic beads Biotinylated Human IL-2 R beta&IL-2 R alpha&IL-2 R gamma Protein, His,&Twin Strep Tag (MALS verified)
ILG-M5253 Mouse Fc Tag Mouse IL-2 R beta&IL-2 R alpha&IL-2 R gamma Protein, Fc Tag&Fc Tag
ILG-C52W3 Cynomolgus His Tag & Strep Tag Cynomolgus IL-2 R beta&IL-2 R alpha&IL-2 R gamma Protein, His Tag&Twin-Strep Tag (MALS verified)
ILG-C82F6 Cynomolgus Fc Tag & Avi Tag Biotinylated Cynomolgus IL-2 R beta&IL-2 R alpha&IL-2 R gamma Protein, Fc,&Fc, (MALS verified)
ILG-R5223 Rat His Tag & Strep Tag Rat IL-2 R beta&IL-2 R alpha&IL-2 R gamma Protein, His Tag&Twin-Strep Tag (MALS verified)
更多IL系列产品
验证数据
亲和力经SPR验证
经SPR验证,IL-2与IL-2受体的亲和力验证结果与文献[4]基本一致

表1 IL-2与LI-2受体结合亲和力数据

IL-2 R

IL-2 R αβγ

IL-2 R βγ

IL-2 R alpha

IL-2 R beta

与Human IL-2亲和力

40.6 pM

0.279 nM

29.9 nM

377 nM

IL-2与IL-2受体蛋白的亲和力验证数据(SPR)

Human IL-2RB&IL-2RA&IL-2RG, Fc Tag&Fc Tag (Cat. No. ILG-H5257) captured on CM5 chip via Anti-human IgG Fc antibodies surface can bind Human IL-2, Tag Free with an affinity constant of 40.6 pM as determined in a SPR assay (Biacore 8K) (Routinely tested).

Protocol

IL-2与IL-2受体蛋白的亲和力验证数据(SPR)

Human IL-2RB&IL-2RG Heterodimer Protein, Fc Tag&Fc Tag (Cat. No. ILG-H5254) captured on CM5 chip via Anti-human IgG Fc antibodies surface can bind Human IL-2, Tag Free with an affinity constant of 0.279 nM as determined in a SPR assay (Biacore 8K) (Routinely tested).

Protocol

IL-2与IL-2受体蛋白的亲和力验证数据(SPR)

Human IL-2 R alpha, His Tag (Cat. No. ILA-H52H9) captured on CM5 chip via anti-His antibody, can bind Human IL-2, Tag Free with an affinity constant of 29.9 nM as determined in a SPR assay (Biacore T200) (Routinely tested).

Protocol

IL-2与IL-2受体蛋白的亲和力验证数据(SPR)

Cynomolgus IL-2 R beta, His Tag (Cat. No. ILB-C52H9) immobilized on CM5 Chip can bind Cynomolgus IL-2, His Tag (Cat. No. IL2-C5249) with an affinity constant of 377 nM as determined in a SPR assay (Biacore 8K) (Routinely tested).

Protocol

高纯度异源聚体经MALS验证
IL-2RB&IL-2RG异源二聚体蛋白(MALS)

The purity of Human IL-2RB&IL-2RG Heterodimer Protein, Fc Tag&Fc Tag (MALS verified)(Cat. No. ILG-H5254) is more than 90% and the molecular weight of this protein is around 145-165 kDa verified by SEC-MALS.

IL-2RB&IL-2RA&IL-2RG异源三聚体蛋白(MALS)

The purity of Human IL-2RB&IL-2RA&IL-2RG, Fc Tag&Fc Tag(Cat. No. ILG-H5257) is more than 90% and the molecular weight of this protein is around 175-190 kDa verified by SEC-MALS.

高生物活性经ELISA验证
IL-2与IL-2受体蛋白的结合验证数据(ELISA)

Immobilized Biotinylated Human IL-2, His,Avitag (Cat. No. IL2-H82E4) at 1 μg/mL (100 μL/well) on streptavidin precoated (0.5 μg/well) plate can bind Human IL-2RB&IL-2RA&IL-2RG, Fc Tag&Fc Tag (Cat. No. ILG-H5257) with a linear range of 0.4-6 ng/mL (QC tested).

Protocol

IL-2与IL-2受体蛋白的结合验证数据(ELISA)

Immobilized Biotinylated Human IL-2, His,Avitag (Cat. No. IL2-H82E4) at 1 μg/mL (100 μL/well) on streptavidin precoated (0.5 μg/well) plate can bind Human IL-2RB&IL-2RA&IL-2RG, Fc Tag&Fc Tag (Cat. No. ILG-H5257) with a linear range of 0.4-6 ng/mL.

Protocol

参考资料:

1. NKTR-2017JPMorganSlides

2. Spolski R, et al. Nat Rev Immunol. 2018. PMID: 30089912 Review.

3. Xinquan Wang et al.,2005, Science. 310(5751):1159-63.

4. Balasubramanian, S. Int Immunol. 1995 Nov;7(11):1839-49.

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