| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
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| 25mg |
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| 500mg |
| Targets |
This compound functions as an ADC linker; the DBCO group binds to azides, the aldehyde binds to aminooxy/hydrazine groups, and the disulfide bond is cleaved by intracellular reducing agents.
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| ln Vitro |
ADCs are made up of an antibody and an ADC cytotoxin that are joined together by an ADC linker[1].
DBCO-SS-aldehyde is a cleavable ADC linker containing DBCO for strain-promoted click chemistry and an aldehyde for oxime ligation, supporting versatile and stable antibody-drug conjugate synthesis. The DBCO group is a strained alkyne that undergoes SPAAC (copper-free click chemistry) with azide-functionalized molecules, offering rapid and bioorthogonal conjugation without the need for a copper catalyst, which is important for preserving the integrity of sensitive biomolecules. The aldehyde group can react with hydrazine or aminooxy groups to form hydrazone or oxime linkages, respectively. The disulfide bond enables selective, controlled release of the payload under reducing conditions (e.g., high intracellular glutathione concentrations). The linker is part of a versatile chemical linker utilized in bioconjugation, enabling the coupling of a drug molecule to an antibody. This strategic linking allows ADCs to selectively target and deliver cytotoxic agents specifically to cancer cells, minimizing harm to healthy cells and enhancing the efficacy of cancer therapy. The linker has no inherent biological activity. It is widely used for the synthesis of ADCs, as well as for protein labeling, surface functionalization, and the development of diagnostic assays. |
| ln Vivo |
As a linker, DBCO-SS-aldehyde is not designed to have direct in vivo activity. The in vivo efficacy of ADCs built with this linker depends on the antibody specificity and payload potency. The disulfide bond provides a cleavable mechanism for intracellular drug release. The SPAAC reaction (DBCO-azide) is highly selective and rapid and can be performed in vivo for pretargeted imaging or therapy applications. The aldehyde group can be used for conjugation to an antibody or other targeting ligand. The disulfide bond is stable in the bloodstream (low glutathione, ~2-20 microM) but is cleaved in the reducing environment of the cell cytoplasm (high glutathione, ~1-10 mM) and the lysosome. This ensures that the payload is released primarily inside target cells. In vivo, ADCs using disulfide linkers have shown efficacy in tumor models. The DBCO group may have some reactivity with biological azides, but the concentration of endogenous azides is very low, so the reaction is highly specific. The linker's in vivo stability and efficacy are optimized for each ADC.
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| Enzyme Assay |
As a chemical linker, DBCO-SS-aldehyde is characterized by chemical assays. The purity (≥95%) is confirmed by HPLC. The structure is verified by ¹H NMR spectroscopy and high-resolution mass spectrometry (HRMS). ¹H NMR (DMSO-d₆) shows: the DBCO aromatic protons as multiplets at delta 7.2-7.8 ppm; the aldehyde proton as a singlet at delta ~9.8-10.0 ppm; the methylene groups as multiplets at delta 2.5-3.5 ppm. The molecular weight (565.66) is confirmed by ESI-MS. The DBCO group content is quantified by reaction with a standard azide (e.g., 4-azidobenzoic acid) in PBS or DMSO at room temperature, monitoring the disappearance of the DBCO peak (UV absorbance at 210-250 nm or by HPLC). The SPAAC reaction is rapid (k2 ~0.1-1 M-¹s-¹). The aldehyde group content is quantified by reaction with a standard hydrazine (e.g., 2,4-dinitrophenylhydrazine, DNPH) under acidic conditions, forming a hydrazone that can be quantified by UV-Vis (absorbance at 360-380 nm for DNPH derivatives). The disulfide bond content is quantified by a thiol release assay: the compound is treated with a reducing agent (e.g., DTT or TCEP) in a buffer (e.g., 0.1 M Tris-HCl, pH 8.0, containing 1 mM EDTA) at 37degC for 30-60 minutes, generating two thiol groups. The thiols are quantified by Ellman's reagent (DTNB), which produces a yellow chromophore (TNB) with absorbance at 412 nm (ε=14,150 M-¹cm-¹).
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| Cell Assay |
As a linker, DBCO-SS-aldehyde is not used directly in cell-based assays. The final conjugate (e.g., ADC, fluorescent probe) is tested. For an ADC built with this linker, a typical cell-based assay involves testing the ADC on antigen-positive and antigen-negative tumor cells. Target cells are seeded in 96-well plates at 5×103 cells per well in medium containing 10% FBS. After overnight attachment, the ADC is added at varying concentrations (typically 0.001-100 nM based on antibody content) and incubated for 72-120 hours. Cell viability is measured by CellTiter-Glo or MTT assay. The IC50 for antigen-positive cells is calculated. To confirm the role of the disulfide bond (cleavable mechanism), the assay may be performed in the presence of a reducing agent such as glutathione (10 mM) to enhance payload release, or an endocytosis inhibitor such as dynasore to block activity. For fluorescence labeling, an azide-functionalized fluorophore is clicked to the DBCO group of the linker, and the resulting conjugate is added to cells. Alternatively, the aldehyde group can be conjugated to an aminooxy probe. Uptake and localization are studied by confocal microscopy.
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| Animal Protocol |
The final conjugate, not the linker alone, is administered to animals. A typical in vivo protocol for an ADC built with DBCO-SS-aldehyde involves a murine xenograft model. Female athymic nude mice are injected subcutaneously with 5-10×10⁶ antigen-positive tumor cells. When tumors reach 100-200 mm3, mice are randomized into treatment groups (n=8-10). The ADC is formulated in PBS and administered intravenously (i.v.) via the tail vein at doses of 1-10 mg/kg (based on antibody content). Treatment is typically administered once weekly for 2-4 weeks. Tumor volumes are measured with calipers every 2-3 days, and body weights are recorded. For pretargeted imaging applications, an antibody-azide conjugate is administered first, allowed to clear, and then the DBCO-SS-aldehyde probe (e.g., DBCO-SS-aldehyde-fluorophore) is administered; the SPAAC reaction occurs in vivo, and the signal is visualized. The disulfide bond can be reduced by tumor microenvironment factors (e.g., glutathione), releasing a payload or imaging agent.
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| ADME/Pharmacokinetics |
The pharmacokinetics (PK) of the final ADC are determined, not the linker alone. For ADCs built with DBCO-SS-aldehyde, the PK is characterized by the antibody portion. The DBCO group is stable in circulation, and the disulfide bond is stable in plasma (low glutathione). The volume of distribution (Vd) is low, similar to plasma volume. Clearance (CL) is low, primarily via catabolism. The half-life is typically 4-7 days for IgG1-based ADCs in mice. The disulfide bond may be reduced by plasma thiols at a low rate, leading to some payload release. The aldehyde group is usually conjugated to an aminooxy/hydrazine group and is not present in the final conjugate as a free aldehyde. For pretargeting applications, the pharmacokinetics of the small molecule probe (e.g., DBCO-SS-aldehyde-fluorophore) are rapid: short half-life (minutes to hours), rapid clearance via the kidneys.
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| Toxicity/Toxicokinetics |
The toxicity of the final ADC, not the linker alone, is evaluated. For ADCs built with disulfide linkers, premature cleavage in circulation can lead to off-target toxicity. The DBCO group has good biocompatibility. For laboratory handling, DBCO-SS-aldehyde should be handled with standard chemical safety precautions: use gloves, lab coat, eye protection. The aldehyde group may be irritating to skin and eyes. Avoid inhalation. The compound is for research use only. Store at -20degC in a dry, dark environment under inert atmosphere to prevent oxidation of the disulfide bond. Avoid co-storage with reducing agents.
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| References |
[1]. Beck A, et al. Strategies and challenges for the next generation of antibody-drug conjugates. Nat Rev Drug Discov. 2017 May;16(5):315-337.
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| Additional Infomation |
DBCO-SS-aldehyde is a cleavable ADC linker used in the synthesis of antibody-drug conjugates (ADCs). It is also a click chemistry reagent containing a DBCO group for copper-free SPAAC and an aldehyde group for oxime/hydrazone ligation. The disulfide bond provides controlled release under reducing conditions. DBCO-SS-aldehyde is a versatile chemical linker utilized for bioconjugation, playing a pivotal role in binding molecules to proteins, peptides, or other biomolecules. Applications include antibody-drug conjugates, protein labeling, click chemistry, and functional biomaterials. It enables precise visualization, tracking, and quantification of proteins in diverse biological studies. The compound is for research use only, not for human therapeutic use. It should be stored at -20degC in a dry environment protected from light.
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| Molecular Formula |
C??H??N?O?S?
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|---|---|
| Molecular Weight |
571.71
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| Appearance |
Typically exists as solid at room temperature
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| Synonyms |
DBCOSSaldehyde; DBCO SS aldehyde
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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 |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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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 | 1.7491 mL | 8.7457 mL | 17.4914 mL | |
| 5 mM | 0.3498 mL | 1.7491 mL | 3.4983 mL | |
| 10 mM | 0.1749 mL | 0.8746 mL | 1.7491 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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