| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Targets |
The biotin moiety targets and binds with extremely high affinity to proteins streptavidin and avidin. The methyltetrazine group is a "bioorthogonal handle" that reacts specifically with TCO. No direct biological target.
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| ln Vitro |
The compound is a bioconjugation reagent. Its primary in vitro activity is the rate of the iEDDA reaction. The methyltetrazine group reacts very rapidly with TCO groups, with second-order rate constants ranging from 1 to 10 M-¹s-¹ or higher. This high speed and specificity enable efficient labeling of TCO-modified biomolecules in complex media, such as cell lysates or on living cell surfaces, without interference from native functional groups.
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| ln Vivo |
This compound is not a drug and has no direct in vivo activity. However, its chemical activity enables a range of in vivo applications. When the TCO-modified targeting vehicle (e.g., an antibody) is pre-injected, a Biotin-PEG4-MeTz probe can be subsequently administered to perform in vivo imaging (e.g., PET or fluorescence) of the target, or to selectively deliver a biotinylated payload. The bioorthogonal reaction proceeds in living animals.
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| Enzyme Assay |
The activity of the methyltetrazine group is confirmed by a non-cellular kinetic assay. A TCO-functionalized fluorescent dye is mixed with Biotin-PEG4-MeTz in a buffer at 37degC. The progress of the click reaction is monitored by HPLC to measure the depletion of the starting materials and the formation of the product. The pseudo-first-order rate constant is calculated based on the change in product concentration over time. This validates the reagent's functionality.
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| Cell Assay |
To demonstrate utility, an in vitro cellular assay for labeling is performed. First, a target cell line is cultured and then treated with a TCO-modified antibody (e.g., a TCO-anti-HER2 antibody). After washing to remove unbound antibody, the cells are incubated with Biotin-PEG4-MeTz. The cells are then washed, and the presence of the biotin label is detected by staining with a fluorescently-labeled streptavidin (e.g., Alexa Fluor 488-streptavidin). A positive fluorescence signal on the cell surface, visualized by flow cytometry or microscopy, confirms successful bioorthogonal labeling.
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| Animal Protocol |
A specific in vivo protocol would involve a two-step approach. Step 1: A mouse with a tumor is intravenously (IV) injected with a TCO-modified tumor-targeting antibody. After a sufficient time for the antibody to accumulate in the tumor (e.g., 24-48 hours), a clearing agent is optionally used to remove unbound antibody from circulation. Step 2: Biotin-PEG4-MeTz, conjugated to a fluorescent or PET imaging agent, is injected IV. The methyltetrazine rapidly reacts with the TCO on the tumor-bound antibody. The animal is then imaged using the appropriate modality (e.g., fluorescence imaging) to visualize the tumor.
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| ADME/Pharmacokinetics |
The pharmacokinetics (PK) of the compound itself are not characterized, as it is not administered alone. Instead, the PK of the final conjugate (e.g., the antibody-biotin-PEG4-MeTz complex) is determined by the carrier molecule. The hydrophilic PEG4 spacer helps to mask the biotin and tetrazine groups, reducing their recognition by the immune system or clearance by the liver, thus potentially prolonging the circulation time of the conjugate.
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| Toxicity/Toxicokinetics |
No specific toxicity information is available for this biotinylated reagent. Biotin and PEG are generally recognized as safe (GRAS) and have low toxicity. Toxicity is not a primary concern for a research reagent used in trace amounts. Standard laboratory safety precautions for handling organic solids should be followed.
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| References |
[1]. Kitowski A, et al. A Sweet Galactose Transfer: Metabolic Oligosaccharide Engineering as a Tool To Study Glycans in Plasmodium Infection. Chembiochem. 2020 Sep 14;21(18):2696-2700.
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| Additional Infomation |
This is a state-of-the-art tool for pretargeted imaging and drug delivery. The strategy aims to improve therapeutic index by decoupling the long time needed for antibody targeting from the fast kinetics of a small molecule radioactive payload. The biotin-streptavidin interaction is one of the strongest known non-covalent bonds, making this an extremely powerful tool for signal amplification in assays. It is strictly a research product.
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| Exact Mass |
674.321
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|---|---|
| CAS # |
1962919-31-8
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| PubChem CID |
164512771
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| Appearance |
Purple to purplish red solid powder
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
12
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| Rotatable Bond Count |
23
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| Heavy Atom Count |
47
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| Complexity |
924
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| Defined Atom Stereocenter Count |
3
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| SMILES |
CC1=NN=C(N=N1)C2=CC=C(C=C2)CNC(=O)CCOCCOCCOCCOCCNC(=O)CCCC[C@H]3[C@@H]4[C@H](CS3)NC(=O)N4
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| InChi Key |
FVPKDWZJCYTWCS-ZEZDXWPOSA-N
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| InChi Code |
InChI=1S/C31H46N8O7S/c1-22-36-38-30(39-37-22)24-8-6-23(7-9-24)20-33-28(41)10-12-43-14-16-45-18-19-46-17-15-44-13-11-32-27(40)5-3-2-4-26-29-25(21-47-26)34-31(42)35-29/h6-9,25-26,29H,2-5,10-21H2,1H3,(H,32,40)(H,33,41)(H2,34,35,42)/t25-,26-,29-/m0/s1
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| Chemical Name |
5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]-N-[2-[2-[2-[2-[3-[[4-(6-methyl-1,2,4,5-tetrazin-3-yl)phenyl]methylamino]-3-oxopropoxy]ethoxy]ethoxy]ethoxy]ethyl]pentanamide
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : 250 mg/mL (370.47 mM)
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| 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
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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 : PEG300 ：Tween 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). View More
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)] 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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
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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