Design and validation of an immuno-PCR assay for IFN-α2b quantification in human plasma

ATTALLAH, CAROLINA; RODRÍGUEZ, MARÍA C; LOZANO, VICTORIA; ETCHEVERRIGARAY, MARINA; OGGERO, MARCOS

Bioanalysis

Future Science

Lugar: London; Año: 2019

1757-6180

Aim: Nowadays, IFN-α is considered a promising therapeutic target for systemic lupus erythematosus. An immuno-PCR (iPCR) was developed to quantify low amounts of IFN-α in human plasma followed by a deep analysis of the methodologic robustness throughout quality by design approach. Results: An accurate, sensitive, selective and versatile iPCR was validated. The critical iPCR procedural steps were identified, applying a Plackett?Burman design. Also, this assay demonstrated an outstanding LOD of 0.3 pg/ml. A significant aspect relies on its high versatility to detect and quantify other cytokines in human plasma as the appropriate biotinylated antibody is employed. Conclusion: This reliable iPCR assay can be clinically used as an alternative method for quantitating and detecting low IFN-α2b concentrations in human plasma samples.