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 |
| Species | Mouse | Rat | Rabbit | Guinea pig | Hamster | Dog |
| Weight (kg) | 0.02 | 0.15 | 1.8 | 0.4 | 0.08 | 10 |
| Body Surface Area (m2) | 0.007 | 0.025 | 0.15 | 0.05 | 0.02 | 0.5 |
| Km factor | 3 | 6 | 12 | 8 | 5 | 20 |
| Animal A (mg/kg) = Animal B (mg/kg) multiplied by | Animal B Km |
| Animal A Km |
For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.
| Related Drug-Linker Conjugates for ADC Products |
|---|
| SC-VC-PAB-MMAE
SC-VC-PAB-MMAE is part of a drug-linker conjugate for ADC (ADC) with anticancer activity, which consists of MMAE (a tubulin inhibitor) linked to a cleavable SC-VC-PAB linker. |
| DBCO-(PEG)3-VC-PAB-MMAE
DBCO-(PEG)3-vc-PAB-MMAE is an active molecule coupler (drug-linker conjugate for ADC) consisting of auristatin E attached to a DBCO-(PEG)3-vc-PAB connector. It can be used for cancer research. In addition, DBCO-(PEG)3-vc-PAB-MMAE is a click chemistry that contains a DBCO moiety and can undergo strain-promoted alkyne-azide cycloaddition (SPAAC) with molecules containing an Azide moiety. |
| PB038
PB038 is a drug-liker containing a PEG unit and a cleavable linker attached to Exatecan. |
| Val-Ala-PAB-MMAE
Val-Ala-PAB-MMAE is a Drug-Linker Conjugate for ADC, consisting of ADC linker (Val-Ala-PAB) and MMAE. |
| Val-Ala-PABC-Exatecan
Val-Ala-PABC-Exatecan is a Drug-Linker Conjugates for ADC, consiting of a cleavable Tesirine linker (Val-Ala-PABC) and Exatecan (topoisomerase I inhibitor). |
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