Avoiding biotin interference in AlphaLISA assays

AlphaLISA™ technology is a highly sensitive, easy-to-use, and reproducible method for detecting and quantifying molecules in various biological matrices. It works by using streptavidin-coated Donor beads and biotinylated anti-analyte antibodies. When these come into close proximity, the excitation of the Donor beads at 680 nM triggers an energy transfer cascade in the Acceptor beads, generating a sharp emission peak at 615 nM.

However, some cell culture media contain high levels of biotin, which can interfere with AlphaLISA and other assay technologies that rely on a streptavidin-biotin binding event for detection. High levels of free biotin in the sample matrix can result in a decrease in total counts, lower signal to background ratios, and reduced AlphaLISA assay detection limits.

To mitigate this, AlphaLISA biotin-free kits have been developed. This application note demonstrates the value of using AlphaLISA biotin-free kits to reduce the effects of biotin interference in sample and standard preparations.

For research use only. Not for use in diagnostic procedures.