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DSS Crosslinker (Disuccinimidyl suberate) is a non-cleavable, cell membrane permeable ADC linker used in the synthesis of antibody-drug conjugates (ADCs). DSS crosslinker has amine-reactive NHS esters at both ends of an 8-atom (11.4 angstrom) spacer arm that are used to conjugate proteins. DSS crosslinker can be used to conjugate or crosslink intracellular proteins.
DSS Crosslinking Protocol
1. Allow vial of DSS Crosslinker to fully equilibrate to ambient temperature before opening to prevent condensation inside the vial (DSS is moisture-sensitive).
2. Prepare a 50 mM solution of DSS, by dissolving 10 mg DSS in 540 µL of dry DMSO or dry DMF.
3. Using a 20-fold excess approach (20:1 Crosslinker:Protein), add crosslinker solution to the protein sample in non-amine containing buffer (i.e. 25 mM Sodium Phosphate, pH 7.4), so that the final crosslinker concentration is between 0.5 to 5 mM. Optimal pH range is from 7 to 9. The DSS solution may become slightly hazy in an aqueous mixture.
4. Allow the sample to react at room temperature for 45 minutes to 1 hour. Allow slightly longer if sample must be kept on ice (recommended 2-3 hours). This reaction rate is not highly temperature sensitive.
5. Quench any unreacted DSS with 25 mM to 200 mM Tris, pH 7.4 and allow to react for 10-15 minutes at room temperature.
6. Desalt sample to remove residual crosslinker (i.e. gel filtration or dialysis, etc.)
| Molecular Weight | 368.34 |
| Formula | C16H20N2O8 |
| CAS Number | 68528-80-3 |
| Solubility (25°C) | DMSO ≥ 90 mg/mL chloroform 50 mg/mL |
| Storage |
Powder -20°C 3 years ; 4°C 2 years In solvent -80°C 6 months ; -20°C 1 month |
| Related Drug-Linker Conjugates for ADC Products |
|---|
| Mc-Val-Cit-PABC-PNP
Mc-Val-Cit-PABC-PNP is a histone-cleavable peptide linker for antibody-drug couplings (ADCs) that can be used for the synthesis of ADCs such as Brentuximab vedotin. |
| DM1-SMCC
DM1-SMCC is DM1 with a reactive linker SMCC, which can react with antibody to make ADC. |
| CL2A-SN-38
CL2A-SN-38 is a SN38 derivative with a peptide-linker which can easily react with antibody to form an ADC. |
| BS3 (bis(sulfosuccinimidyl)suberate)
BS3 (bis(sulfosuccinimidyl)suberate) is a non-cleavable ADC linker used in the synthesis of antibody-drug conjugates (ADCs). |
| Sulfo-SMCC sodium
Sulfo-SMCC sodium is a commonly used hetero-bifunctional, noncleavable ADC crosslinker bearing N-hydroxysuccinimide (NHS) ester and maleimide groups to react with primary amines and sulfhydryl groups, respectively. |
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