Active Mannose Associated Serine Protease 2 (MASP2)

MASP2 (Mannan-binding lectin serine protease 2) is a serum protease that plays an important role in the activation of the complement system via mannose-binding lectin. The preproprotein of MASP2 is proteolytically processed to generate A and B chains that heterodimerize to form the mature protease, which is able to associate with MBL2. Thus, a functional binding ELISA assay was constructed to detect the association of rhMASP2 with MBL2. Briefly, rhMASP-2 were diluted serially in 10mM Tris-HCl, 1M NaCl, 5mM CaCl2, and 0.05%Triton X-100 (pH 7.4). Duplicate samples of 100uL were then transferred to MBL2-coated microtiter wells and incubated for 2h at 37oC. Wells were washed with PBST and incubated for 1h with anti-MASP-2 pAb, then aspirated and washed 3 times. After incubation with HRP labeled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution , wells were incubated for 15-25 minutes at 37oC. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of MASP2 with MBL2 was shown in Figure 1 and this effect was in a dose dependent manner.