Ramucirumab (anti-VEGFR2)

Catalog No.A2003 Batch: A200301

Technical Data

CAS No.	947687-13-0
Formulation	PBS buffer, pH 7.2
Isotype	human IgG1
Storage (From the date of receipt)	Store the undiluted solution at 4°C in the dark to avoid freeze-thaw cycles
Purity	99.4%
Protein concentration	5mg/ml
Endotoxin Level	≤1 EU/mg

Biological Activity

Description

Ramucirumab is a monoclonal antibody of the IgG1 class that binds to VEGF-R2 and prevents its activation. The IC50 value for blocking KDR binding to VEGF is 0.8 nM for ramucirumab, MW: 143.6 KD.

Targets

KDR/VEGF interaction ^[5]

0.8 nM

In vitro

Ramucirumab is direct inhibitor of VEGF-R2, where it binds to the extracellular VEGF-binding domain with high degree of specificity and affinity, at picomolar dose range. It thus prevents the binding of the VEGF ligand to the VEGF-R2 receptor. Ramucirumab has potential advantages over bevacizumab as it is selective for VEGF-R2, whereas bevacizumab by targeting VEGF-A affects VEGF-R1, -R2, and the noncatalytic coreceptors neuropilin-1and -2^[1]. Ramucirumab prevents the binding of VEGFR ligands: VEGF-A, VEGF-C, and VEGF-D to its receptors. Thus, ramucirumab inhibits the angiogenesis pathways involved in the development and progression of gastric cancer^[2]. ramucirumab demonstrates inhibition of VEGF-stimulated VEGFR-2 activation, proliferation of human endothelial cells, VEGF migration of human leukemia cells, and VEGF induced phosphorylation of VEGFR-2 in human umbilical vein and porcine aortic endothelial cells overexpressing VEGFR-2^[3]. Preclinical in vitro data revealed that ramucirumab has a high affinity for VEGFR-2, showing a half-maximal effective concentration (EC50) of 0.15 nM, 8- to 9-fold higher than its natural ligand VEGFA. Ramucirumab binds VEGFR-2 in domain 3 near the N-terminus, both as a soluble protein and as a cell-surface receptor, with a IC50 of 1-2 nM^[4].

In vivo

In preclinical studies, ramucirumab concentrations of >20 μg/mL were associated with anticancer activity^[1]. And ramucirumab potently inhibited VEGF binding to VEGFR-2 with a binding affinity constant of ramucirumab to VEGFR-2 of 5 × 10^-11 M. Pharmacokinetic evaluation has demonstrated a nonlinear pharmacokinetic, with incremental doses of this agent being associated with a decrease in clearance. Ramucirumab has a half-life of 200-300 h^[3]. Preclinical studies were also to be conducted in mice, but interspecies receptor differences made ramucirumab inactive in preclinical mouse models^[4].

Protocol (Only for Reference)

Cell Assay:	The anti-KDR antibodies were added to the microwells of a 96-well plate coated with KDR(Ig1-7) (full-length KDR ECD), KDR(Ig1-3) (variant that contains only the first three N-terminal Ig domains of KDR ECD), or KDR(Ig1) (variant that contains only the first N-terminal Ig domain of KDR ECD) and incubated at room temperature for 1 h. The plate was washed and then incubated with a mouse anti-human Fc antibody-HRP conjugate for additional 1 h, after which the plate was developed as described above. Antibody binding to the two Ig deletion variants are shown as relative (as percentages) to their binding to the full-length KDR ECD.

Cell Assay:

The anti-KDR antibodies were added to the microwells of a 96-well plate coated with KDR(Ig1-7) (full-length KDR ECD), KDR(Ig1-3) (variant that contains only the first three N-terminal Ig domains of KDR ECD), or KDR(Ig1) (variant that contains only the first N-terminal Ig domain of KDR ECD) and incubated at room temperature for 1 h. The plate was washed and then incubated with a mouse anti-human Fc antibody-HRP conjugate for additional 1 h, after which the plate was developed as described above. Antibody binding to the two Ig deletion variants are shown as relative (as percentages) to their binding to the full-length KDR ECD.

References	[1] Javle M, et al. Clin Cancer Res. 2014, 20(23):5875-81. [2] Oholendt AL, et al. J Adv Pract Oncol. 2015, 6(1):71-5. [3] Spratlin JL, et al. Future Oncol. 2010, 6(7):1085-94. [4] Diaz-Serrano A, et al. Expert Rev Anticancer Ther. 2016, 16(6):585-95. [5] Lu D, et al. J Biol Chem. 2003, 278(44):43496-507. + Expand

References

+ Expand

Selleck's Ramucirumab (anti-VEGFR2) has been cited by 2 publications

[ Int J Mol Sci, 2024, ]	PubMed: 38339049
Analysis of Tumor Heterogeneity Through AXL Activation in Primary Resistance to EGFR Tyrosine Kinase Inhibitors [ JTO Clin Res Rep, 2023, 4(6):100525]	PubMed: 37426308

FOR RESEARCH USE ONLY. NOT FOR USE IN HUMANS.

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