Strandh M, Persson B, Roos H, Ohlson S
Department of Natural Sciences, University of Kalmar, Sweden.
J Mol Recognit. 1998 Winter;11(1-6):188-90. doi: 10.1002/(SICI)1099-1352(199812)11:1/6<188::AID-JMR420>3.0.CO;2-O.
Interactions between the immobilized weak-affinity monoclonal IgG antibody 39.5, which is specific for the glucose-alpha 1,4-glucose motif, and various oligosaccharides were studied with surface plasmon resonance technology. The antibody was immobilized at high levels on the surface of the sensor chip and different concentrations of the analytes were injected at 25 and 40 degrees C. The 39.5 antibody exhibited specific binding to maltose, tetraglucose and maltotriose, with dissociation constants Kd in the range from 0.07 mM (25 degrees C) to 1.0 mM (40 degrees C). Association and dissociation rate constants (ka and kd) were rapid and baseline was obtained almost immediately after the end of each antigen injection. This excluded the need for a regeneration step but also made calculation of the kinetic values impossible. Owing to the weak affinity and the small size of the analytes (< 1000 Da), a careful design of control surfaces is demanded to exclude artefactual results.
利用表面等离子体共振技术研究了固定化的对葡萄糖-α-1,4-葡萄糖基序具有特异性的弱亲和力单克隆IgG抗体39.5与各种寡糖之间的相互作用。该抗体以高水平固定在传感器芯片表面，并在25℃和40℃下注入不同浓度的分析物。39.5抗体对麦芽糖、四糖和麦芽三糖表现出特异性结合，解离常数Kd在0.07 mM（25℃）至1.0 mM（40℃）范围内。缔合和解离速率常数（ka和kd）很快，每次抗原注射结束后几乎立即获得基线。这排除了再生步骤的需要，但也使得动力学值的计算变得不可能。由于亲和力较弱且分析物尺寸较小（<1000 Da），需要精心设计对照表面以排除人为结果。
