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| 1mg |
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| 5mg |
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| 10mg |
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| Targets |
M2e, human TFA targets the immune system, specifically inducing antibodies that bind to the extracellular domain of the influenza A virus M2 ion channel protein expressed on the surface of infected cells, leading to antibody-dependent cell-mediated cytotoxicity (ADCC).
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| ln Vitro |
M2e, human TFA acts as an effective and versatile vaccine candidate against human influenza A strains, including both seasonal and pandemic subtypes. As a synthetic peptide, it can induce cross-protective immune responses when appropriately formulated with adjuvants.
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| ln Vivo |
In vivo animal studies demonstrate that M2e-based vaccines can elicit protective immune responses against heterologous influenza A virus challenges. When administered intranasally or parenterally with appropriate adjuvants, M2e induces antibodies that reduce viral load and protect against morbidity and mortality.
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| Enzyme Assay |
Non-cell binding assays for M2e peptide are typically performed to evaluate its immunogenicity or to detect anti-M2e antibodies. For an ELISA to detect anti-M2e antibodies, 96-well plates are coated with 1-5 microg/mL of M2e, human TFA in PBS and incubated overnight at 4degC. The plates are blocked with 3% BSA or 5% non-fat milk for 2 hours at room temperature, then washed with PBS-T (0.05% Tween-20). Serially diluted serum samples (from immunized animals) are added to the plates and incubated for 2 hours. After washing, HRP-conjugated anti-mouse (or anti-human) IgG is added, followed by TMB substrate. The reaction is stopped with H2SO4, and absorbance is measured at 450 nm. For peptide-MHC binding assays, computational methods or in vitro refolding assays with purified MHC class II molecules are used. Alternatively, Surface Plasmon Resonance (SPR) can be performed with M2e-specific monoclonal antibodies to determine binding affinity.
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| Cell Assay |
Cellular assays for M2e, human TFA are performed using primary immune cells (splenocytes, PBMCs) harvested from M2e-immunized mice or humans. For T cell recall responses, 5×10⁵ splenocytes are seeded in 96-well plates and stimulated with 1-50 microg/mL M2e, human TFA for 48-72 hours. Cell culture supernatants are collected, and cytokine production (IFN-gamma, IL-4, IL-5, IL-10) is measured by ELISA. For T cell proliferation, cells are labeled with CFSE (5 microM) prior to M2e stimulation, and proliferation (CFSE dilution) is analyzed by flow cytometry at 72-96 hours. For B cell ELISpot, 96-well PVDF plates are coated with anti-mouse Ig capture antibody, and splenocytes are added with M2e peptide. After 16-24 hours, antibody-secreting cells are detected with HRP-labeled detection antibody and AEC substrate, and spots are counted using an automated ELISpot reader. For intracellular cytokine staining (ICS), cells are stimulated with M2e peptide plus Brefeldin A for 4-6 hours, then stained for surface markers (CD4, CD8) and intracellular cytokines (IFN-gamma, TNF-alpha, IL-2) after fixation and permeabilization, followed by flow cytometry.
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| Animal Protocol |
For in vivo immunization studies, 6-8 week old female BALB/c or C57BL/6 mice are immunized with M2e, human TFA formulated with an adjuvant (e.g., Freund's adjuvant, alum, or more advanced adjuvants like CpG ODN or AddaVax). Mice receive 10-50 microg of M2e peptide per dose, administered via intraperitoneal, subcutaneous, or intranasal routes, typically in 2-3 doses at 2-4 week intervals. Blood samples are collected via retro-orbital bleeding at baseline and 10-14 days after each immunization to assess serum antibody titers by ELISA. For challenge studies, 4-6 weeks after the final immunization, mice are anesthetized with isoflurane and infected intranasally with a lethal dose (e.g., 1-10× LD50) of influenza A virus (e.g., H1N1, H3N2, or avian H5N1). Body weight and survival are monitored daily for 14-21 days. At 3-7 days post-infection, mice are euthanized for viral load determination. Lung tissues are homogenized in PBS and clarified by centrifugation, and viral titers are determined by plaque assay using MDCK cells or by quantitative RT-PCR (qRT-PCR) for influenza M gene. Lung samples are also fixed, sectioned, and stained with H&E for histopathological analysis of inflammation and lung damage.
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| ADME/Pharmacokinetics |
As a peptide-based immunogen, M2e, human TFA (MW ~2738.83 Da) is not a classic drug and its pharmacokinetic properties are not extensively characterized. When administered in vivo with adjuvant, the peptide remains in the injection site (depot effect) for days to weeks, depending on the adjuvant formulation. The peptide is taken up by antigen-presenting cells (dendritic cells, macrophages) at the injection site via phagocytosis and macropinocytosis. After internalization, M2e is processed and presented via MHC class II to CD4+ T cells and cross-presented via MHC class I to CD8+ T cells. The TFA salt form improves solubility for formulation. Systemic absorption of intact peptide is minimal. The elimination half-life is not relevant due to its immunogen mechanism of action.
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| Toxicity/Toxicokinetics |
Specific toxicity data for M2e, human TFA is limited as it is a research peptide. In animal immunization studies using M2e peptide with adjuvants, the toxicity profile is primarily determined by the adjuvant rather than the peptide itself. At typical immunization doses (10-50 microg per mouse), M2e peptide is generally well-tolerated, with no overt signs of systemic toxicity. Local injection site reactions (mild swelling, redness) may occur due to adjuvant components. Long-term safety studies are not available due to the compound's research status. The TFA counterion may cause irritation at high doses.
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| References | |
| Additional Infomation |
M2e (matrix protein 2 ectodomain) is highly conserved across human influenza A virus strains, making it a key target for developing a "universal" influenza vaccine that would provide protection against multiple strains without annual reformulation. M2e, human TFA is used as a research tool in the development of such vaccines. Its mechanism of action is based on eliciting antibodies that bind to the M2 protein on infected cells, leading to elimination via ADCC. The peptide has not yet received regulatory approval for human use, but is widely studied in preclinical and early-phase clinical trials. The TFA salt form enhances solubility and stability for research use.
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| Molecular Formula |
C109H171F3N32O43S2
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| Molecular Weight |
2738.83
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| Related CAS # |
M2e, human;251359-24-7
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| Appearance |
White to off-white solid powder
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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, avoid exposure to moisture. |
| 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 :~100 mg/mL (~36.51 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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.3651 mL | 1.8256 mL | 3.6512 mL | |
| 5 mM | 0.0730 mL | 0.3651 mL | 0.7302 mL | |
| 10 mM | 0.0365 mL | 0.1826 mL | 0.3651 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
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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