BEVACIZUMAB

Cat No.:V4373 Purity: ≥98%
Bevacizumab (trade nameAvastin; Anti-Human VEGF, Humanized Antibody) is a humanized monoclonal antibody which binds specifically to allVEGF-Aisoforms with high affinity.
BEVACIZUMAB Chemical Structure CAS No.: 216974-75-3
Product category: Others 8
This product is for research use only, not for human use. We do not sell to patients.
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1mg
5mg
10mg
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Other Forms of BEVACIZUMAB:

  • Bevacizumab (Avastin)
Official Supplier of:
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Top Publications Citing lnvivochem Products
Product Description

Bevacizumab (trade name Avastin; Anti-Human VEGF, Humanized Antibody) is a humanized monoclonal antibody which binds specifically to all VEGF-A isoforms with high affinity. It is a medication used to treat a number of types of cancers and a specific eye disease. For cancer it is given by slow injection into a vein and used for colon cancer, lung cancer, glioblastoma, and renal-cell carcinoma. For age-related macular degeneration it is given by injection into the eye.

Biological Activity I Assay Protocols (From Reference)
ln Vitro
A humanized monoclonal antibody called bevacizumab selectively and highly binds to all VEGF-A isoforms, blocking their interaction with VEGFR-1 and VEGFR-2[1]. According to experimental investigation, bevacizumab's EC50 for binding VEGF, as determined by ELISA, is 0.18 μg/mL. Similar findings from binding kinetics experiments indicate that bevacizumab suppresses HUVEC proliferation driven by VEGF, with an IC50 value of 0.047±0.0081 μg/mL[2].
ln Vivo
In comparison to the control group, it has been shown that the subconjunctival administration of FD006 and Bevacizumab can significantly decrease CoNV in NaOH cauterized rats (p < 0.01)[2].
Animal Protocol
Kinase Assay: The binding kinetics of Bevacizumab or FD006 to VEGF is measured using Bio-Layer Inter-Ferometry on Octet RED. The assay is conducted at 30°C in PBS buffer. Sensor tips are pre-wet for 15 mins in buffer immediately prior to use, and the microplates are filled with 200 μL per well of diluted samples (VEGF) or buffer and agitated at 1000 rpm. The anti-human IgG biosensor are pre-saturated with Bevacizumab or FD006 (10 μg/mL) and washed in buffer for 120 seconds, and then transferred to VEGF at concentrations of 10 μg/mL, 3 μg/mL and 1 μg/mL. The VEGF association and dissociation rates are measured for 5mins and 10mins, respectively. The Kinetics parameters (Kon and Koff) and affinities (KD) are calculated from a non-linear global fit using the Octet analysis software. Multiple independent measurements are performed.



Cell Assay: Human umbilical vein endothelial cells (HUVECs) (1×104 cells/100 μL/well) are seeded in 96-well plates and cultured at 37 for 14 h with Endothelial Cell Medium supplemented with 5% heat-inactivated FCS, 100 U/mL Penicillin, 100 U/mL Streptomycin, and endothelial cell growth supplement. After low-serum starvation overnight, cells are treated with different concentrations of FD006 or Bevacizumab which are pre-incubated with 10 ng/mL VEGF for 30 minutes and incubated at 37, 5% CO2 for 72 hours. Then, 10 μL CCK8 is added to each well and incubated for another 4 hours. The absorbance is measured by spectrophotometer at 450 nm to determine the cell viability.


Animal Studies: After modeling, ninety rats are randomly divided into five groups (eighteen rats per group) and receive a subconjunctival injection with 0.05 mL per rat of (1) 0.9% NaCl, (2) solvent, (3) 0.1% Dexamethasone (Dexamethasone sodium phosphate), (4) 25 mg/mL Bevacizumab and (5) 25 mg/mL FD006 in the superior temporal conjunctiva on the day after modeling. All chemical burns and treatments are performed by one investigator. The operator is blinded to the treatment group from which each cornea is derived. At postoperative days 3, 7, 14, 21 and 28, the eyes are harvested for further studies after the rats are sacrificed with an overdose of 10% chloral hydrate.

References
[1]. Tan H, et al. 99mTc-labeled bevacizumab for detecting atherosclerotic plaque linked to plaque neovascularization and monitoring antiangiogenic effects of treatment in ApoE-/-mice. Sci Rep. 2017 Jun 14;7(1):3504.
[2]. Wang Q, et al. Pharmacological characteristics and efficacy of a novel anti-angiogenic antibody FD006 in corneal neovascularization. BMC Biotechnol. 2014 Feb 27;14:17.
[3]. Di Mauro C, et al. Hedgehog signalling pathway orchestrates angiogenesis in triple-negative breast cancers. Br J Cancer. 2017 May 23;116(11):1425-1435
These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Weight
0
CAS #
216974-75-3
Related CAS #
Bevacizumab (PBS);216974-75-3
Storage

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Shipping Condition
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
Solubility Data
Solubility (In Vivo)
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

Injection Formulations
(e.g. IP/IV/IM/SC)
Injection Formulation 1: DMSO : Tween 80: Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution 50 μL Tween 80 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO 400 μLPEG300 50 μL Tween 80 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).
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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C,1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO 100 μLPEG300 200 μL castor oil 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10: 10 : 80 (i.e. 100 μL Ethanol 100 μL Cremophor 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH 400 μLPEG300 50 μL Tween 80 450 μL Saline)


Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).
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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders


Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

 (Please use freshly prepared in vivo formulations for optimal results.)
Calculator

Molarity Calculator allows you to calculate the mass, volume, and/or concentration required for a solution, as detailed below:

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An example of molarity calculation using the molarity calculator is shown below:
What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
  • Enter 350.26 in the Molecular Weight (MW) box
  • Enter 10 in the Concentration box and choose the correct unit (mM)
  • Enter 5 in the Volume box and choose the correct unit (mL)
  • Click the “Calculate” button
  • The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

Dilution Calculator allows you to calculate how to dilute a stock solution of known concentrations. For example, you may Enter C1, C2 & V2 to calculate V1, as detailed below:

What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
  • Enter 10 into the Concentration (Start) box and choose the correct unit (mM)
  • Enter 25 into the Concentration (End) box and select the correct unit (mM)
  • Enter 25 into the Volume (End) box and choose the correct unit (mL)
  • Click the “Calculate” button
  • The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box
g/mol

Molecular Weight Calculator allows you to calculate the molar mass and elemental composition of a compound, as detailed below:

Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
Instructions to calculate molar mass (molecular weight) of a chemical compound:
  • To calculate molar mass of a chemical compound, please enter the chemical/molecular formula and click the “Calculate’ button.
Definitions of molecular mass, molecular weight, molar mass and molar weight:
  • Molecular mass (or molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
  • Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.
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Reconstitution Calculator allows you to calculate the volume of solvent required to reconstitute your vial.

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  • The answer appears in the Volume (to add to vial) box
In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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Calculation results

Working concentration mg/mL;

Method for preparing DMSO stock solution mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.

Method for preparing in vivo formulation:Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.

(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
             (2) Be sure to add the solvent(s) in order.

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