Biotin and Streptavidin/Avidin

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Biotin is highly hydrophobic and only sparingly water-soluble (0.19 mg/mL; saturated solution measured at CellMosaic®). Most commercially available biotinylation kits therefore require a high percentage of organic solvent to dissolve biotin or activated biotin. Organic solvents can induce antibody aggregation and promote precipitation. Based on our experience, antibodies and proteins labeled with biotin can aggregate and precipitate from solution over time. Historically, to optimize performance, we have limited biotin labeling to 2–4 biotins per antibody/protein. However, in many applications, higher biotin loading is desirable to enable signal amplification or improved selection.

AqT® linkers are super-hydrophilic, water-soluble, charge-neutral proprietary materials invented at CellMosaic®. We have modified biotin with an AqT® linker to increase biotin’s hydrophilicity and make it water-soluble, enabling biotinylation with minimal or no organic solvent. AqT-biotin forms a network of hydrogen bonds (H-bonds) with surrounding water via AqT hydroxyl (–OH) groups, creating a microenvironment that helps prevent neighboring biotins from stacking or interacting with one another. As a result, antibodies can be labeled with higher levels of AqT-biotin without altering their properties. This AqT-enhanced H-bond network may also help protect antibodies from enzymatic degradation and support retention of activity.

CellMosaic commercializes AqT® biotin labeling kits and products. If you are interested in co-developing AqT® biotin-based products, click here to learn about partnering with CellMosaic. For a detailed description of the AqT® technologies, please see our technology section.

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