Strohner Pavel, Korn Stephanie, Buhl Roland, Becher Gunther
BioTeZ Berlin-Buch, Berlin, Germany.
J Immunoassay Immunochem. 2013;34(1):83-93. doi: 10.1080/15321819.2012.683501.
The therapeutic use of antibodies has grown exponentially, providing new treatment options for various diseases. Monitoring treatment with therapeutic antibodies is a particular challenge because often the target antigen is no longer measurable or the results are unreliable. To overcome this problem, a recovery-ELISA was developed to quantify therapeutic antibody and antigen by a modification of the traditional sandwich immunoassay using omalizumab as an example. Standard serum samples were spiked with IgE and omalizumab in a certain concentration range to create standard curves. After incubation and washout procedures, the reaction was stopped and the plate read with bichromatic absorbance. This approach defines reference functions for serum IgE without and with antibody, respectively. Based on these functions, free IgE and omalizumab concentrations can be calculated in serum samples of patients treated with omalizumab. The recovery-ELISA allows us to easily quantify antigen (free IgE) and therapeutic antibody (omalizumab) simultaneously in the same sample and thus to monitor patients treated with the specific antibody.
抗体的治疗用途呈指数级增长，为各种疾病提供了新的治疗选择。监测治疗性抗体的治疗是一项特殊挑战，因为通常靶抗原不再可测量或结果不可靠。为克服这一问题，以奥马珠单抗为例，通过对传统夹心免疫测定法进行改进，开发了一种回收率酶联免疫吸附测定法（recovery-ELISA）来定量治疗性抗体和抗原。将标准血清样本与一定浓度范围内的IgE和奥马珠单抗混合，以创建标准曲线。经过孵育和洗脱程序后，终止反应，并用双色吸光度读取平板。该方法分别定义了无抗体和有抗体时血清IgE的参考函数。基于这些函数，可以计算接受奥马珠单抗治疗患者血清样本中的游离IgE和奥马珠单抗浓度。回收率酶联免疫吸附测定法使我们能够在同一样本中轻松同时定量抗原（游离IgE）和治疗性抗体（奥马珠单抗），从而监测接受特定抗体治疗的患者。
