CD47蛋白——肿瘤免疫治疗热门靶点

背景

产品列表

验证数据

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
CD47	CD7-H5227	Human	Human CD47 Protein, His Tag^HOT
	CD7-H5256	Human	Human CD47 Protein, Fc Tag (HPLC-verified)^HOT
	CD7-H82E9	Human	Biotinylated Human CD47 Protein, His,Avitag™^HOT
	CD7-H82F6	Human	Biotinylated Human CD47 Protein, Fc,Avitag™^HOT
	CD7-H52A5	Human	Human CD47 Protein, Mouse IgG2a Fc Tag, low endotoxin^HOT
	EP-102	Human	CD47: SIRP alpha [Biotinylated] Inhibitor Screening ELISA Assay Pair^HOT
	CD7-H522a	Human	Human CD47 Protein, His Tag, low endotoxin
	CD7-H5251	Human	Human CD47 Protein, Llama IgG2b Fc Tag, low endotoxin
	CD7-H82F8	Human	Biotinylated Human CD47 Protein, Fc (L234A, L235A, P329G),Avitag™
	CD7-HA2E9	Human	Unconjugated Human CD47 Protein, His,Avitag™
	CD7-M522b	Mouse	Mouse CD47 Protein, His Tag
	CD7-M5251	Mouse	Mouse CD47 Protein, Fc Tag
	CD7-M525a	Mouse	Mouse CD47 Protein, Mouse IgG2a Fc Tag, low endotoxin
	CD7-M82E4	Mouse	Biotinylated Mouse CD47 Protein, His,Avitag™
	CD7-C5252	Cynomolgus / Rhesus macaque	Cynomolgus / Rhesus macaque CD47 Protein, Fc Tag
	CD7-C52H1	Cynomolgus / Rhesus macaque	Cynomolgus / Rhesus macaque CD47 Protein, His Tag
	CD7-R5256	Rat	Rat CD47 Protein, Fc Tag
	CD7-R5257	Rabbit	Rabbit CD47 Protein, Fc Tag
	CD7-C52H5	Canine	Canine CD47 Protein, His Tag
SIRP alpha	CDA-H82F2	Human	Biotinylated Human SIRP alpha / CD172a Protein, Fc,Avitag™ (MALS verified)
	SIA-H5225	Human	Human SIRP alpha / CD172a Protein, His Tag (MALS verified)
	SIA-H5251	Human	Human SIRP alpha / CD172a Protein, Fc Tag (MALS verified)
	SIA-H52A8	Human	Human SIRP alpha / CD172a Protein, Mouse IgG1 Fc Tag (MALS verified)
	SIA-H82E0	Human	Biotinylated Human SIRP alpha / CD172a Protein, His,Avitag™ (MALS verified)
	SIA-HP252	Human	PE-Labeled Human SIRP alpha / CD172a Protein, Fc Tag (recommended for neutralizing assay)
	SIA-M5252	Mouse	Mouse SIRP alpha / CD172a Protein, Mouse IgG2a Fc Tag, low endotoxin
	SIA-M5258	Mouse	Mouse SIRP alpha / CD172a Protein, Fc Tag
	SIA-M52H4	Mouse	Mouse SIRP alpha / CD172a Protein, His Tag
	SIA-C5254	Cynomolgus	Cynomolgus SIRP alpha / CD172a Protein, Fc Tag
	SIA-C52H7	Cynomolgus	Cynomolgus SIRP alpha / CD172a Protein, His Tag
SIRP beta	SIA-H5257	Human	Human SIRP beta / CD172b Protein, Fc Tag
SIRP gamma	SIG-H5253	Human	Human 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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