Why Choosing the Right Validation Method is Important
Detection and quantification of protein isoforms by ELISA is challenging and requires careful validation and choice of assay antibodies. Mature BDNF is such an example, as BDNF ELISA assays often cross-react with the full-length proBDNF isoform, or actual isoform preference that has not been tested. Additionally, choosing an incorrect immunological technique to validate isoform detection may lead to false conclusions.
Polacchini et al., 2015 and Satori et al., 2019, reported that the Biosensis Mature BDNF RapidTM ELISA quantifies 'total BDNF', based on line blot experiments using the assay's detection antibody. We have therefore re-assessed our kit performance, and compared it to a competitor BDNF ELISA (Company A), claimed to be mature BDNF-specific in the very same publications.
The Figure above is an example from our Technical Note #5 showing ~4-Fold higher molar cross-reactivity of proBDNF proteins from 5 different sources in Company A's Mature BDNF ELISA (Red), as compared to the Biosensis Mature BDNF RapidTM ELISA assay (Black).
Our Technical Note #5 summarises our findings and confirms our initial claims of preferential quantification of mature BDNF with our Mature BDNF RapidTM ELISA.
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