【靶点聚焦】VEGF，横跨癌症、眼科、心血管疾病等多种疾病的泛用性靶点

血管内皮生长因子（VascularEndothelial Growth Factor,VEGF）是一种高度特异性的促血管内皮细胞生长因子，在胚胎发育、骨骼生长和生殖等生理过程中的对血管生成起关键调节作用，同时与肿瘤相关的病理性血管生成有关。

VEGF信号系统较为复杂，包含5种配体(VEGF-A、VEGF-B、VEGF-C、VEGF-D和PIGF)及三种酪氨酸激酶受体(VEGFR1、VEGFR2和VEGFR3)，各配体对不同受体的亲和力和选择性存在差异。在所有受体之中VEGFR-2扮演着最主要的作用，几乎介导所有VEGF已知的细胞应答。VEGFA与VEGFR2受体结合诱导的信号传导是血管生成的主要控制者，会激活细胞内信号通路从而刺激肿瘤血管内皮细胞的生长、增殖和成熟及新生血管生成。

• VEGF与癌症

目前，对于VEGF促进肿瘤血管生成的作用及其与人类癌症发病机制的关系已经有较为明确的研究成果。大多数恶性肿瘤中都能观察到VEGF及其mRNA高表达，特别在肿瘤组织血管增生丰富的部位，肿瘤细胞和周围基质分泌的VEGF会刺激内皮细胞的增殖和存活，导致新血管的形成，新血管可能结构异常和渗漏，并与侵袭性、血管密度、转移、复发和预后相关。因此靶向VEGF是癌症治疗的的一种潜在方式，许多小分子抑制剂也是基于该信号通路开发。

• VEGF与眼科疾病

临床上有很多新生血管性眼病都是因为眼内VEGF过表达使得新生血管生长，继而发生大量出血、纤维增殖、牵拉性视网膜脱离、新生血管性青光眼等严重并发症。争性抑制VEGF-R2，能够有效抑制血管生成并促进已有新生血管消退，减轻血管渗漏引起的渗出、水肿和炎性反应，从而减缓眼底新生血管的进展。在眼科，使用抑制VEGF药物有助于阻断病变新生血管的生长，从而治疗眼科疾病。

为进一步满足VEGF相关研究需求，加速相应癌症及眼科疾病等治疗药物开发，ACROBiosystems基于对其信号传导机制的研究，在原有的VEGF系列靶点蛋白产品及磁珠产品基础上，最新开发了VEGFR报告基因细胞系，致力于为客户提供从蛋白到细胞株的全系列配套产品，从药物筛选到细胞功能性验证等不同场景全面加速相关抗体药及小分子抑制剂开发进程。

VEGF系列蛋白

点击分子查看详细信息及验证数据

中和实验表明，抗VEGF抗体能抑制VEGF165蛋白(Cat. No. VE5-H4210)对HUVEC细胞的促增殖作用，ED50值为0.065-0.229 ug/ml。

Neutralization assay shows that the proliferation effect of ActiveMax® Human VEGF165 (Cat. No. VE5-H4210) was inhibited by increasing concentration of anti-VEGF mAb. The concentration of VEGF165 used is 20 ng/ml. The ED50 is 0.065-0.229 ug/ml (Routinely tested).

VEGFR报告基因细胞系

VEGFR2(Luc) HEK293 Reporter Cell (Cat. No.CHEK-ATF044)是在HEK293细胞内过表达VEGFR2的同时转入一个受NFAT信号调控转录的Luciferase报告基因。游离的VEGF与该细胞膜表面的VEGFR2结合后，激活NFAT信号，诱导Luciferase报告基因表达。

VEGFR2(Luc) HEK293 Reporter Cell (Cat. No.CHEK-ATF044)可用于筛选VEGF/VEGFR途径的阻断抗体或小分子等药物。

1.细胞性质验证

经FACS及Signaling Bioassay验证，Cat.No. CHEK-ATF044表达VEGFR，用连续稀释的VEGF165(Cat.No. VE5-H4210)进行刺激，EC50值为5.13ng/mL，最大诱导倍数约为70。

Expression analysis of human VEGFR2 on VEGFR2 (Luc) HEK293 Reporter Cell by FACS. Cell surface staining was performed on VEGFR2 (Luc) HEK293 Reporter Cellor negative control cell using PE-labeled anti-VEGFR2 antibody.

Response to human VEGF165 protein (RLU).The VEGFR2 (Luc) HEK293 Reporter Cell was stimulated with serialdilutions of human VEGF165 protein (Cat. No. VE5-H4210).The EC50 was approximately 5.13 ng/mL.

The VEGFR2 (Luc) HEK293 Reporter Cell was stimulated with serialdilutions of human VEGF165 protein (Cat. No. VE5-H4210).The max induction fold was approximately 70.

2.代次稳定性验证

通过FACS及Signaling Bioassay对Cat.No. CHEK-ATF044传代稳定性进行验证，均证明该细胞株可稳定传递10-32代。

Passage stability analysis of receptor expression by FACS. Flow cytometry surface staining of human VEGFR2 on VEGFR2 (Luc)HEK293 Reporter Cell demonstrates consistent mean fluorescent intensity across passage10-32.

Passage stability analysis by Signaling Bioassay. The continuously growing VEGFR2 (Luc) HEK293 Reporter Cell was stimulated with serial dilutions of human VEGF165 protein. Human VEGF165 protein stimulated response demonstrates passage stabilization (fold induction and EC50) across passage 10-32.

1.筛选中和抗体

经验证，Bavacizumab与抗VEGF中和抗体均可以抑制VEGF蛋白(Cat.No. VE5-H4210)诱导的报告基因活性，EC50值为分别为0.0071 μg/mL；0.015 μg/mL。

Inhibition of human VEGF165 protein-induced reporter activity by anti-human VEGF neutralizing antibody.This reporter cell was incubated with serial dilutions of antibodiesin the presence of human VEGF165 protein (Cat.No.VE5-H4210)with a final concentration of 10 ng/mL. EC50 values were as follows: Bavacizumab 0.0071 μg/mL, Human VEGF165 Neutralizing Antibody 0.015 μg/mL. 

2.筛选小分子抑制剂

经验证，VEGFR小分子抑制剂可以抑制VEGF蛋白(Cat.No. VE5-H4210)诱导的报告基因活性，EC50值为0.0053μM。

Inhibition of human VEGF165 protein-induced reporter activity by human VEGFR2 small molecule inhibitor. This reporter cell was incubated with serial dilutions of inhibitors inthe presence of human VEGF165 protein (Cat. No. VE5-H4210)with a final concentration of 10 ng/mL. The EC50 of human VEGFR2 small molecule inhibitor (Cabozantinib) was approximately 0.0053 μM.

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VEGF 预偶联磁珠

为满足VEGF相关抗体筛选需求，ACROBiosystems还开发了人源VEGF165预偶联磁珠（Cat.No.MBS-K036），可作为高效低成本的抗体筛选工具，欢迎点击了解。