Development and characterization of monoclonal antibodies against inhibin α subunit as a potential cancer biomarker

Inhibin is a dimeric glycoprotein hormone consisting of α and β subunits. Inhibin has gained significant interest as a biomarker for ovarian and testicular malignancies. While existing inhibin immunoassays have demonstrated efficacy, assays specifically targeting the free α subunit are not yet available. This study aimed to generate monoclonal antibodies against inhibin-α using hybridoma technology, targeting distinct epitopes within the N-region of the α subunit. These epitopes were chosen using bioinformatic analysis to ensure strong antigenicity and successful antibody production. To screen the antibodies, an ELISA was developed using plates coated with recombinant inhibin-α protein. This ELISA confirmed that the antibodies were reactive and could effectively detect inhibin-α in vitro. The antibodies were further evaluated for specificity and immunoreactivity using Western blotting, and IHC methods. Western blot analysis showed that the antibodies specifically recognized the free inhibin-α, demonstrating high specificity under both reducing and non-reducing conditions. IHC studies on normal mouse tissue sections further confirmed the localization of inhibin-α in granulosa, theca, Sertoli, and Leydig cells. The developed antibodies showed significant potential for use in vitro assays for cancer diagnosis. Further validation is needed to ensure their successful clinical application.