Poly-N-Isopropylacrylamide Hydrogel Based Fluoroimmunoassay of Methyltestosterone

A novel immunoassay method for methyltestosterone (MT) in serum ovas developed.The antigen of MT (Ag(MT))was synthesized by covalently bonding MT to BSA and raising its antibody (Ab(MT)). Ab(MT) was con.jugated to poly-N-isopropylacrylamide (p-NIPAAm) to form Ab(MT) bound thermally reversible hydrogel p-NIPAAm-Ab((MT) A competitive immunoassay method based on the competition of fluorescein isothiocyanate (FITC) labeled MT antigen (FITC-Ag(MT)) and free MT with limited amount of p-NIPAAm-Ab(MT) was established. The separation in the assay process was achieved by precipitation of the immuno-complex above its critical solution temperature. The detection limit for MT is 50ng/ml. The recoveries of MT from human serum are satisfactory.