Phospholipase B (PLB) is a multifunctional enzyme that exhibits both phospholipase A1/A2 and lysophospholipase activities, enabling it to hydrolyze phospholipids at both the sn-1 and sn-2 positions. Primarily found in fungi, bacteria, and mammalian tissues, PLB plays critical roles in lipid metabolism, membrane remodeling, and signal transduction. In pathogenic fungi like Candida albicans, PLB contributes to virulence by facilitating host cell invasion and immune evasion.Besides,Phospholipase C Beta 2 (PLCb2) has been identified as an interactor of PLB, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human PLB and recombinant human PLCb2. Briefly, PLB was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to PLCb2-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-PLB pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630nm immediately. The binding activity of recombinant human PLB and recombinant human PLCb2 was shown in Figure 1, the EC50 for this effect is 1.72 µg/mL.