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CD47蛋白——肿瘤免疫治疗热门靶点

背景
CD47,也被称作整合素相关蛋白(Integrin-associated protein, IAP),是所有细胞表达的靶点中“marker of self”的典型代表。CD47是组织稳态的关键调节器, 同时也在从动脉粥样硬化到癌症的多种疾病中发挥作用。CD47通过逃逸吞噬作用来保护宿主细胞,癌细胞表达的CD47与NK细胞表达的SIRP家族蛋白SIRPα以及SIRPγ相互作用。通过这样的方式保护癌细胞避免被吞噬消除。
另外,CD47对于TSP-1介导的血管NO信号通路也是必不可少的。它通过调节CD36和VEGFR2以抑制血管生成。 NO信号通路的长期失调也是目前几种心血管疾病的标志,因此CD47成为动脉粥样硬化的药物靶点。目前研究表明,在多种体内模型中抗CD47单抗可减轻动脉粥样硬化程度。
由于CD47在癌症和心血管疾病的发病机理中的重要作用, CD47信号通路已被广泛研究用于新的靶向疗法的开发。国内外多家药企布局该靶点抗体药物的研发。 ACROBiosystems提供了不同种属以及多种标签类型的CD47&SIRPα重组蛋白(包括生物素标记的重组蛋白)以及对应的抗体筛选试剂盒。该系列产品可用于抗体的免疫制备,中和抗体的快速高通量筛选,细胞学验证以及种属交叉反应等多个应用场景。 满足ELISA, SPR, BLI, FACS, Alphalisa和MSD等多个实验平台。
产品列表
分子名 货号 种属 产品描述
CD47CD7-H5227HumanHuman CD47 Protein, His TagHOT
CD7-H5256HumanHuman CD47 Protein, Fc Tag (HPLC-verified)HOT
CD7-H82E9HumanBiotinylated Human CD47 Protein, His,Avitag™HOT
CD7-H82F6HumanBiotinylated Human CD47 Protein, Fc,Avitag™HOT
CD7-H52A5HumanHuman CD47 Protein, Mouse IgG2a Fc Tag, low endotoxinHOT
EP-102HumanCD47: SIRP alpha [Biotinylated] Inhibitor Screening ELISA Assay PairHOT
CD7-H522aHumanHuman CD47 Protein, His Tag, low endotoxin
CD7-H5251HumanHuman CD47 Protein, Llama IgG2b Fc Tag, low endotoxin
CD7-H82F8HumanBiotinylated Human CD47 Protein, Fc (L234A, L235A, P329G),Avitag™
CD7-HA2E9HumanUnconjugated Human CD47 Protein, His,Avitag™
CD7-M522bMouseMouse CD47 Protein, His Tag
CD7-M5251MouseMouse CD47 Protein, Fc Tag
CD7-M525aMouseMouse CD47 Protein, Mouse IgG2a Fc Tag, low endotoxin
CD7-M82E4MouseBiotinylated Mouse CD47 Protein, His,Avitag™
CD7-C5252Cynomolgus / Rhesus macaqueCynomolgus / Rhesus macaque CD47 Protein, Fc Tag
CD7-C52H1Cynomolgus / Rhesus macaqueCynomolgus / Rhesus macaque CD47 Protein, His Tag
CD7-R5256RatRat CD47 Protein, Fc Tag
CD7-R5257RabbitRabbit CD47 Protein, Fc Tag
CD7-C52H5CanineCanine CD47 Protein, His Tag
SIRP alphaCDA-H82F2HumanBiotinylated Human SIRP alpha / CD172a Protein, Fc,Avitag™ (MALS verified)
SIA-H5225HumanHuman SIRP alpha / CD172a Protein, His Tag (MALS verified)
SIA-H5251HumanHuman SIRP alpha / CD172a Protein, Fc Tag (MALS verified)
SIA-H52A8HumanHuman SIRP alpha / CD172a Protein, Mouse IgG1 Fc Tag (MALS verified)
SIA-H82E0HumanBiotinylated Human SIRP alpha / CD172a Protein, His,Avitag™ (MALS verified)
SIA-HP252HumanPE-Labeled Human SIRP alpha / CD172a Protein, Fc Tag (recommended for neutralizing assay)
SIA-M5252MouseMouse SIRP alpha / CD172a Protein, Mouse IgG2a Fc Tag, low endotoxin
SIA-M5258MouseMouse SIRP alpha / CD172a Protein, Fc Tag
SIA-M52H4MouseMouse SIRP alpha / CD172a Protein, His Tag
SIA-C5254CynomolgusCynomolgus SIRP alpha / CD172a Protein, Fc Tag
SIA-C52H7CynomolgusCynomolgus SIRP alpha / CD172a Protein, His Tag
SIRP betaSIA-H5257HumanHuman SIRP beta / CD172b Protein, Fc Tag
SIRP gammaSIG-H5253HumanHuman SIRP gamma / CD172g Protein, Fc Tag
验证数据
高均一性——HPLC验证
Fig1. The purity of Human CD47, Fc Tag (HPLC-verified) (Cat. No. CD7-H5256) was greater than 90% as determined by SEC-HPLC.

Protocol

CD47与SIRPα的结合验证
> ELISA验证CD47蛋白与SIRP-a结合

Fig2.Immobilized Human SIRP alpha, His Tag (HPLC-verified) (Cat. No. SIA-H5225) at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD47, Fc,Avitag (Cat. No. CD7-H82F6) with a linear range of 8-250 ng/mL.

Protocol

> BLI验证CD47蛋白与SIRP-a结合

Fig3.Immobilized Human CD47, Fc Tag (HPLC-verified) (Cat. No. CD7-H5256) on AHC Biosensor, can bind Human SIRP alpha, His Tag (HPLC-verified) (Cat. No. SIA-H5225) with an affinity constant of 0.72 μM as determined in BLI assay (ForteBio Octet Red96e)

Protocol

> FACS验证,CD47蛋白可与表达SIRP-a的ACHN细胞结合

Fig4.FACS assay shows that Human CD47, Fc Tag (HPLC-verified) (Cat. No. CD7-H5256) can bind to ACHN cell expressing SIRP-a. The concentration of CD47 used is 10 μg/mL.

Protocol

CD47的抗体筛选验证
抗体亲和力测试
> ELISA验证,CD47与抗体结合亲和力强
Fig5.Immobilized Anti-CD47 MAb, Human IgG4 at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD47, His,Avitag (Cat. No. CD7-H82E9) with a linear range of 0.05-2 ng/mL

Protocol

> SPR/BLI验证,CD47与抗体结合亲和力强

Fig6.Anti-Human CD47 MAb (Human IgG4) captured on CM5 chip via Anti-Human IgG Fc antibodies surface, can bind Human CD47, His Tag (Cat. No. CD7-H5227) with an affinity constant of 1.66 nM as determined in a SPR assay (Biacore T200)

Protocol

Fig7.Loaded Anti-Human CD47 MAb (Human IgG4) on AHC Biosensor, can bind Human CD47, His Tag (Cat. No. CD7-H5227) with an affinity constant of 6.18 nM as determined in BLI assay (ForteBio Octet Red96e)

Protocol

中和抗体的筛选策略

Fig8.Serial dilutions of Anti-Human CD47 Neutralizing Antibody were added into Human SIRP alpha, His Tag (HPLC-verified) (Cat. No. SIA-H5225): Biotinylated Human CD47, Fc,Avitag (Cat. No. CD7-H82F6) binding reactions. The half maximal inhibitory concentration (IC50) is 124.5 ng/mL.

Protocol

Fig9.FACS analysis shows that the binding of Human CD47 to ACHN expressing SIRP-a was inhibited by increasing concentration of neutralizing SIRP-a antibody with the IC50 15.02 μg/mL. The concentration of Human CD47 used is 10 μg/mL.

Protocol

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