Effect of trifluoroethanol on antibodies binding properties

S. Bobrovnik, M. O. Demchenko, S. Komisarenko
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Abstract

The studies on the influence of organic co-solvents on the structure and function of antibodies are of key interest, especially in view of antibodies broad use as recognizing elements in different analytical systems. Here we studied the effect of co-solvent 2,2,2-trifluoroethanol (TFE) on the ability of anti-ovalbumin monoclonal antibodies to interact with its specific antigen. Antibody affinity to antigen and the rate constants of antibody binding to immobilized antigen were analyzed. Changes in antibody reactivity with incubation time which depended on TFE concentration and temperature were revealed. When treatment of antibodies with TFE was carried out at 0°C, we observed nonlinear, non-monotonous changes of antibody reactivity with initial fast decrease and substantial increase that may be related to the loss of antigen binding reactivity by some part of antibodies at the start but its restoration when the incubation proceeds. Keywords: 2;2;2-trifluoroethanol, antibody affinity, antigen-antibody interaction, monoclonal antibodies, ovalbumin
三氟乙醇对抗体结合特性的影响
有机共溶剂对抗体结构和功能的影响的研究具有重要意义,特别是考虑到抗体在不同的分析系统中广泛用作识别元件。本文研究了共溶剂2,2,2-三氟乙醇(TFE)对抗卵清蛋白单克隆抗体与其特异性抗原相互作用能力的影响。分析了抗体对抗原的亲和力和抗体与固定化抗原的结合速率常数。抗体反应性随孵育时间的变化取决于TFE浓度和温度。在0℃条件下用TFE处理抗体时,我们观察到抗体反应性的非线性、非单调变化,从最初的快速下降到大幅上升,这可能与抗体的一部分在开始时失去抗原结合反应性,但在孵育过程中恢复有关。关键词:2;2;2-三氟乙醇;抗体亲和;抗原-抗体相互作用
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