Kynurenine 3-monooxygenase (KMO), also known as kynurenine 3-hydroxylase, is an enzyme that plays a crucial role in the metabolism of kynurenine, a derivative of tryptophan, an essential amino acid. is an enzyme that plays a crucial role in the metabolism of kynurenine, a derivative of tryptophan, an essential amino acid. The study revealed that the combination of Aminoadipate Aminotransferase (AADAT) with KMO can affect its catalytic activity and metabolic pathway, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant humant KMO and recombinant human AADAT. Briefly, Briefly, biotin-linked KMO were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to AADAT-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µl stop solution to the wells and read at 450nm immediately. The binding activity of KMO and AADAT was shown in Figure 1, the EC50 for this effect is 0.22ug/mL.