Bevacizumab

Bevacizumab是一种人源化的anti-VEGF单克隆抗体，是VEGF的抑制剂。可与所有人源VEGF-A亚型（和具有生物活性的蛋白水解片段）结合、中和。MW:149 KD。
批次： A200601纯度：98.94 %蛋白浓度：5.0 mg/ml内毒素：≤1 EU/mg

生物活性：产品描述    Bevacizumab是一种人源化的anti-VEGF单克隆抗体，是VEGF的抑制剂。可与所有人源VEGF-A亚型（和具有生物活性的蛋白水解片段）结合、中和。MW:149 KD。
靶点    
VEGF [1]
()
体外研究    
人源VEGF的Gly88对bevacizumab的结合是基本必需的，同时也保证了bevacizumab结合的种属特异性，大鼠和小鼠的VEGF在该对应位置为丝氨酸残基。bevacizumab可与所有人源VEGF-A亚型（和具有生物活性的蛋白水解片段）结合、中和，而不中和其他VEGF基因家族成员，如VEGF-B或VEGF-C[1]。

体内研究    在人体中，bevacizumab的终末半衰期为17-21天。在裸鼠中，Bevacizumab抑制人源肿瘤细胞系的生长，在剂量为1-2 mg/kg(一周给药两次)时，达到最大抑制作用。半最大抑制作用所需剂量为0.1-0.5 mg/kg。在Macaca fascicularis (cynomolgus monkey)中对bevacizumab进行安全性评估研究，对其进行bevacizumab给药4-13周后，年轻成年的食蟹猴骨骺发育不良：肥大的软骨细胞剂量依赖式增多，生长区血管浸润受到抑制。长期bevacizumab的给药抑制了雌性系统的血管生长、卵巢和子宫的体重下降，黄体缺失。生长板和卵巢的改变为可逆性改变，停止治疗可逐渐恢复。除此以外，在药物浓度高达50 mg/kg，也没有其他与给药相关的副作用被观察到[1]。皮下注射后，rhuMAb VEGF（bevacizumab）在大鼠中的生物利用度为69%，而在小鼠和食蟹猴中为100%。Bevacizumab与灵长类VEGF结合，与兔子VEGF低亲和力结合，而不与大鼠、小鼠VEGF结合[2]。
细胞实验：
Objective: Effects of bevacizumab on VEGF-induced HUVEC proliferation
Cells: Human umbilical vein endothelial cells (HUVECs)
Concentrations: 0~500 ng/mL
Incubation Time: 96 h
Method: 50 ng/ml of rhVEGF was pre-incubated with a concentration range of bevacizumab (0-500 ng/ml) for 2 h before added to HUVACs. The plate was continuously incubated for 4 days. The proliferation of cells was determined by using Alamar Blue dye.
Reference: https://www.ncbi.nlm.nih.gov/pubmed/15886877

Objective: Cell proliferation assay
Cells: non small cell lung cancer (NSCLC), colorectal cancer (CRC), breast cancer (BC) and renal cell carcinoma (RCC)
Concentrations: 250 μg/ml
Incubation Time: 24, 48, 72 or 96 hours
Method: Between 2 × 103 and 5 × 103 cells/well (cell line/doubling time dependent) are seeded into 96 well plates and incubated overnight to adhere. Medium is then replaced by RPMI-1640 with reduced FBS and bevacizumab or VEGFA at the concentrations indicated (time point zero). After 24, 48, 72 or 96 hours in hypoxia, MTT (5 mg/ml in PBS) is added and incubated for 2 hours at 37°C. The supernatant is removed and reaction products are solubilized for 1 h in 10% HCl, 0.1% NP-40 in isopropanol. Absorbance is measured at 570 nm with a reference wavelength of 650 nm using an ELISA reader. Each experimental condition is analyzed in triplicate and results are an average of a minimum of three biological repetitions.
Reference: https://www.ncbi.nlm.nih.gov/pubmed/24059699

Bevacizumab can apply to nude mice, various cancer cell lines and other related assays (Only for Reference)
动物实验:    - Collapse
Objective: To determine the antitumor response of bevacizumab and cisplatin as single agents or in combination in xenograft models of ovarian cancer
Animal Models: Nude mice were injected (i.p.) with 5×106 A2780 cells
Formulation: PBS
Dosages: 5mg/kg
Administration: i.p.
Reference: https://www.ncbi.nlm.nih.gov/pubmed/19047105

Objective: To determine the pharmacokinetics of bevacizumab
Animal Models: Female nude mice
Formulation: --
Dosages: 9.3 mg/kg
Administration: i.v. or s.c. injection
Reference: https://www.ncbi.nlm.nih.gov/pubmed/9862791