本研究采用表面等离子体共振技术,以血管紧张素转换酶(angiotensin I converting enzyme,ACE)为蛋白配体,分析马氏珠母贝肉蛋白酶解产物(protein hydrolysate of Pinctada martensii,PHPM)超滤组分与配体的结合情况,利用质谱鉴定结合肽...本研究采用表面等离子体共振技术,以血管紧张素转换酶(angiotensin I converting enzyme,ACE)为蛋白配体,分析马氏珠母贝肉蛋白酶解产物(protein hydrolysate of Pinctada martensii,PHPM)超滤组分与配体的结合情况,利用质谱鉴定结合肽段的氨基酸序列后,筛选潜在抑制ACE活性强的肽段进行合成,研究其体外ACE抑制活性、抑制类型及多肽与ACE蛋白的相互作用。结果显示,PHPM分子质量在3 000~5 000 Da的超滤组分与ACE蛋白具有较强的结合信号,在结合物质的肽序列中优选出4种具有潜在活性的ACE抑制肽进行合成,其中多肽SLPWPMKPMNLIE的半数抑制浓度最低,并且通过氢键与ACE蛋白C端结构域的疏水口袋结合。展开更多
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.
