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.