Quantifying cell binding kinetics mediated by surface-bound blood type B antigen to immobilized antibodies  

作　　者：LI BaoXia CHEN Juan LONG Mian 

机构地区：[1]National Microgravity Laboratory and Center of Biomechanics and Bioengineering, Institute of Mechanics, Chinese Academy ofSciences, Beijing 100190, China

出　　处：《Chinese Science Bulletin》2008年第23期3634-3641,共8页

基　　金：the High-Tech Research and Development Program of China (Grant No. 2007AA02Z306);National Basic Research Program of China (Grant No. 2006CB910303);National Natural Science Foundation of China (Grant Nos. 30730032, 10332060, 30225027)

摘　　要：Cell adhesion is crucial to many biological processes, such as inflammatory responses, tumor metastasis and thrombosis formation. Recently a commercial surface plasmon resonance (SPR)-based BIAcore biosensor has been extended to determine cell binding mediated by surface-bound bio-molecular interactions. How such cell binding is quantitatively governed by kinetic rates and regulating factors, however, has been poorly understood. Here we developed a novel assay to determine the binding kinetics of surface-bound biomolecular interactions using a commercial BIAcore 3000 bio-sensor. Human red blood cells (RBCs) presenting blood group B antigen and CM5 chip bearing immobilized anti-B monoclonal antibody (mAb) were used to obtain the time courses of response unit, or sensorgrams, when flowing RBCs over the chip surface. A cellular kinetic model was proposed to correlate the sensorgrams with kinetic rates. Impacts of regulating factors, such as cell concentration, flow duration and rate, antibody-presenting level, as well as pH value and osmotic pressure of suspending medium were tested systematically, which imparted the confidence that the approach can be applied to kinetic measurements of cell adhesion mediated by surface-bound biomolecular interactions. These results provided a new insight into quantifying cell binding using a commercial SPR-based BIAcore biosensor.Cell adhesion is crucial to many biological processes, such as inflammatory responses, tumor metastasis and thrombosis formation. Recently a commercial surface plasmon resonance （SPR）-based BIAcore biosensor has been extended to determine cell binding mediated by surface-bound bio- molecular interactions. How such cell binding is quantitatively governed by kinetic rates and regulating factors, however, has been poorly understood. Here we developed a novel assay to determine the binding kinetics of surface-bound biomolecular interactions using a commercial BIAcore 3000 bio- sensor. Human red blood cells （RBCs） presenting blood group B antigen and CM5 chip bearing immo- bilized anti-B monoclonal antibody （mAb） were used to obtain the time courses of response unit, or sensorgrams, when flowing RBCs over the chip surface. A cellular kinetic model was proposed to correlate the sensorgrams with kinetic rates. Impacts of regulating factors, such as cell concentration, flow duration and rate, antibody-presenting level, as well as pH value and osmotic pressure of sus- pending medium were tested systematically, which imparted the confidence that the approach can be applied to kinetic measurements of cell adhesion mediated by surface-bound biomolecular interactions. These results provided a new insight into quantifying cell binding using a commercial SPR-based BIAcore biosensor.

关 键 词：生物传感器 细胞动力模式 细胞学 构成模式 

分 类 号：Q2[生物学—细胞生物学]