The influence of naturally occurring heterophilic anti-immunoglobulin antibodies on direct measurement of serum proteins using sandwich ELISAs
The influence of naturally occurring heterophilic anti-immunoglobulin antibodies on direct measurement of serum proteins using sandwich ELISAs
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DOI：
10.1016/s0022-1759(99)00206-9
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发表时间：
2000-02-21
影响因子：
2.2
通讯作者：
Kirchner, H
中科院分区：
文献类型：
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作者：
Hennig, C;Rink, L;Kirchner, H
夹心ELISA已成为一种广泛使用的血清蛋白定量检测方法。然而，它们可能会受到各种干扰物质的影响。正如最近报道的，我们观察到假阳性水平的干扰素（IFN）-α和-β在高达27%的健康献血者使用商业ELISA血清。我们现在证明，两组不同的天然存在的嗜异性抗体（IgG型）是负责这些滴度。组I（代表85%的阳性样品）结合来自山羊、小鼠、大鼠、马和牛科动物（但不结合兔）的IgG的Fab区。组II（15%）识别小鼠、马、牛和兔（但不包括山羊或大鼠）免疫球蛋白Fc区的表位。抗体不与人IgG亚类交叉反应，但有助于使用市售ELISA获得的假阳性IgG类风湿因子水平。为了研究测定对这些伪影的敏感性，已经测试了捕获和检测抗体的各种组合。在此基础上，我们定义了标准ELISA可能受嗜异性抗免疫球蛋白抗体影响的相对风险。一般而言，使用单克隆抗体进行捕获和检测的测定比包括至少一种多克隆抗血清的其它测定更不敏感。然而，已经发现只有利用兔F（ab '）（2）片段的系统对这种干扰免疫。(C)2000爱思唯尔科技有限公司。保留所有权利。
Sandwich ELISAs have become a widely used method for the quantitative detection of serum proteins. However, they can be biased by a variety of interfering substances. As reported recently, we observed false-positive levels of interferon (IFN)-alpha and -beta in up to 27% of sera from healthy blood donors using commercial ELISAs. We now demonstrate that two different groups of naturally occurring heterophilic antibodies (IgG-type) are responsible for these titers. Group I (representing 85% of positive samples) binds to the Fab region of IgG from goat, mouse, rat, horse, and bovidae (but not rabbit). Group II (15%) recognizes an epitope in the Fc region of mouse, horse, bovine, and rabbit (but not goat or rat) immunoglobulins. The antibodies did not crossreact with human IgG subclasses but contributed to false-positive IgG rheumatoid factor levels obtained using a commercially available ELISA. To investigate the susceptibility of assays to these artifacts, various combinations of capture and detection antibodies have been tested. On this basis, we defined the relative risks that standard ELISAs might be influenced by heterophilic anti-immunoglobulin antibodies. In general, assays that use monoclonal antibodies for both capture and detection are less susceptible than others which include at least one polyclonal antiserum. However, only systems utilizing rabbit F(ab')(2) fragments have been found to be immune to this interference. (C) 2000 Elsevier Science B.V. All rights reserved.