The neutralization of interferons by antibody III. The constant antibody bioassay, a highly sensitive quantitative detector of low antibody levels.
The neutralization of interferons by antibody III. The constant antibody bioassay, a highly sensitive quantitative detector of low antibody levels.
复制标题
抗体III对干扰素的中和。
DOI：
10.1089/jir.2008.0043
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Grossberg,LeslieD
中科院分区：
文献类型：
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作者：
Grossberg,SidneyE;Kawade,Yoshimi;Grossberg,LeslieD
在干扰素（IFN）治疗期间，患者体内产生的中和抗体（nab）会降低其有益作用。目前普遍采用的测定NAb的方法是恒定ifn法，将单一给定浓度的抗原与连续稀释的血清混合，最低的最终稀释度（通常为1:20）受到人血清对培养中的人细胞潜在不利影响的限制。本文描述的恒定抗体（Ab）方法使用一定稀释度（通常为1:20）的血清与一系列IFN浓度混合。基于先前提出的Ab-IFN反应模型的理论中和曲线在此以实验可观察到的量来描述。实验结果表明，恒Ab法可将Ab的检测下限提高10-20倍。在理论预测和实验结果之间观察到的极好的一致性加强了使用基于IFN活性降低10倍的效价作为NAb单位的有效性，如先前推荐的那样，报告为10倍还原单位(TRU)/mL。对接受Rebif或Betaseron治疗的患者先前认为阴性的血清（恒ifn法<20 TRU/mL）进行恒抗体检测显示，约50%的患者可检测到nab；avonex治疗患者的血清滴度<1 TRU/mL。恒Abmethod可作为定量的、灵敏的任何性质的IFN NAb筛选生物测定，并且应该能够在IFN治疗过程的早期检测低水平的NAb。该方法可用于检测单克隆抗体，否则无法检测到的nab。原则上，恒Abmethod应适用于任何细胞因子或其他蛋白质效应分子的抗体测定。
The neutralizing antibodies (NAbs) that develop in patients during interferon (IFN) therapy can reduce its beneficial effects. The universally employed method of NAb measurement currently is theconstant IFNmethod, in which antigen at a single given concentration is mixed with serial dilutions of serum, the lowest final dilution of which (usually 1:20) is constrained by the potential adverse effect of human serum on human cells in culture. Theconstant antibody (Ab)method described herein uses serum at a certain set dilution (usually 1:20) mixed with a series of IFN concentrations. Theoretical neutralization curves based on the previously presented model of the Ab-IFN reaction are depicted herein in terms of experimentally observable quantities. As predicted by the theoretical studies, theconstant Abmethod was demonstrated experimentally to extend the lower limits of detection of Ab by a factor of 10–20. The excellent agreement observed between the theoretical prediction and experimental findings reinforces the validity of using as NAb unitage the titer based on 10-fold reduction of IFN activity, reportable as Tenfold Reduction Units (TRU)/mL, as previously recommended. Testing by theconstant Abmethod of sera previously considered negative (<20 TRU/mL by theconstant IFNmethod) from patients treated with Rebif or Betaseron showed that ∼50% had detectable NAbs; such sera from Avonex-treated patients had titers of <1 TRU/mL. Theconstant Abmethod can be used as a quantitative, sensitive IFN NAb screening bioassay of any nature, and should be able to detect low levels of NAbs early in the course of IFN therapy. The method may be useful to test monoclonal antibodies for otherwise undetectable NAbs. In principle, theconstant Abmethod should be applicable to the measurement of NAbs against any cytokine or other protein-effector molecule.