The kinetic analysis of the interaction between tumor necrosis factor(TNF) and its monoclonal antibody was performed by surface plasmon resonance(SPR) technique. The monoclonal antibody was immobilized to the surface ...The kinetic analysis of the interaction between tumor necrosis factor(TNF) and its monoclonal antibody was performed by surface plasmon resonance(SPR) technique. The monoclonal antibody was immobilized to the surface of CM5 sensor chip by amine coupling. TNF at different concentrations was injected across the mAb immobilized surface. The interaction was recorded in real time and could be seen on the sensorgram. One cycle, including association, dissociation and regeneration, lasted no more than 15 min . The interaction results was evaluated using 1∶1 Langmuir binding model. The kinetic rate constants were calculated to be: k a=1.68×10 3 L·mol -1 ·s -1 , k d=1.73×10 -4 s -1 , and the affinity constants K A=9.7×10 6 L·mol -1 , K D=1.03×10 -7 mol·L -1 . The χ \+2 was 3.47, which showed that the interaction is consistent with the 1∶1 model. We can see from the results that although there are two binding sites in one mAb molecule, TNF reacts with each site in an independent and noncooperative manner.展开更多
文摘The kinetic analysis of the interaction between tumor necrosis factor(TNF) and its monoclonal antibody was performed by surface plasmon resonance(SPR) technique. The monoclonal antibody was immobilized to the surface of CM5 sensor chip by amine coupling. TNF at different concentrations was injected across the mAb immobilized surface. The interaction was recorded in real time and could be seen on the sensorgram. One cycle, including association, dissociation and regeneration, lasted no more than 15 min . The interaction results was evaluated using 1∶1 Langmuir binding model. The kinetic rate constants were calculated to be: k a=1.68×10 3 L·mol -1 ·s -1 , k d=1.73×10 -4 s -1 , and the affinity constants K A=9.7×10 6 L·mol -1 , K D=1.03×10 -7 mol·L -1 . The χ \+2 was 3.47, which showed that the interaction is consistent with the 1∶1 model. We can see from the results that although there are two binding sites in one mAb molecule, TNF reacts with each site in an independent and noncooperative manner.
