alpha-ketoglutarate (alpha-ICA) can convert to 2-hydroxygiutarate (24-HG), which is confitmecr to be associated with many diseases, especially with acute myeloid leukemia (AML). In this paper, a novel reaction-based chemosensor DT based on the typical Schiff-base reaction was designed for sensing the biomarker of alpha-KA, in which a diazanyl group as the recognition group was-linked with a benzothiadiazole unit as the fluorophore moiety. Considering the typical Scbiff-base reaction to generate hydrazones suffering from slow kinetics, particularly under neutral conditions,,a series Of parallel-exp: eriments was canducted for optimizing the chemical recognition process, including varying the solvent, -reaction temperature, reactant concentration, and reaction rate. The optimum condition was established as a pH value; temperature, alpha-ICA:concentration, and response time of 5.7, 30 degrees C; 100 mu M, and 20 min, respectively. Notably, in contrast withthe initial 6:3-fold fluorescence enharidement, the remarkable 75-fold fluorescence enhancement ((I - I-0)/I-0 at 560 nm) was observed by-optimizing the chemical recognition process of DT and alpha-ICA Finally, DT was carried out for the chemical recognition, processing of alpha-KA in serum, We demonstrated that DT is selective for alpha-KA over other potential biologically interferences with similar structures and thus is suitable for detecting alpha-ICA in serum. On the basis of the Optimized chemical recognition process, DT shows high potential application for sensing alpha-KA with remarkable fluorescence enhancement. This work provided a potential Method that is quick and convenient for sensing biomarker alpha-KA in serum. It is worth rioting that without complicated pretreatment, utilizing a novel reaction-based fluorescent chemosensor may establish a new promising platform for clinical diagnosis Biomarker: