The S-4FB (succinimidyl-4-formylbenzamide) heterobifunctional crosslinker is fundamental to Chromalink™ technology. S-SS-4FB is an analog of S-4FB with a cleavable disulfide moiety in the linker enabling intracellular cleavage of the conjugate. S-SS-4FB analog reacts with primary amines on proteins (amino group of lysine), amino-modified oligonucleotides or surfaces, introducing a 4FB (4-formylbenzamide) linker that forms stable covalent conjugates with biomolecules possessing HyNic (6-hydrazinonicotinamide) incorporated linkers. 4FB groups incorporated on biomolecules and surfaces are extremely stable and have been known to maintain their reactivity for >18 months. Intracellularly, disulfide bonds of S-SS-4FB are cleaved by glutathione cleaving a drug payload from its carrier.
Chromalink™ technology is based on the use of two complementary heterobifunctional linkers:
- S-HyNic crosslinker or its analog is conjugated to biomolecule 1 through primary amines on proteins, oligos, peptides, or surfaces.
- S-4FB (succinimidyl-4-formylbenzamide) linker or its analog is conjugated to biomolecule 2 through primary amines on proteins, oligos, peptides, carbohydrates, or surfaces.
- HyNic-modified biomolecule 1 is incubated with 4FB-modified biomolecule 2 to form a conjugate.
The result is two biomolecules conjugated through a UV-traceable and stable bond (bis-arylhydrazone) with measurable absorbance at 354 nm. Thus any two proteins, regardless of molecular weight, can be efficiently conjugated.
- Catalyzed Conjugation – Faster kinetics for greater efficiency and yields
- Quantifiable – Conjugate bond is UV-traceable for simple and direct quantification
- Stability – 10 times more stable than any other conjugation linker
- Specificity – Two-linker method avoids homoconjugate formation
- Efficient – >80% efficient linker-biomolecule conjugations
- Robust - The conjugate bond is stable to 92˚C and to pH values ranging from pH 2.0–10.0
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