Anja Schmidt, Diana Stichel, Emilia Salzmann-Manrique, Christoph Königs
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引用次数: 0
摘要
在常规治疗过程中,经常要对 A 型血友病或获得性 A 型血友病患者的血浆样本进行 FVIII 特异性抗体分析。贝塞斯达检测法只能检测抑制性抗体,而酶联免疫吸附法可以检测抑制性和非抑制性抗体。然而,患者血浆样本中经常含有内源性或替代的 FVIII,因此会干扰这两种类型的分析。热变性是灭活 FVIII 的一种方法,但不幸的是,热变性会导致高背景信号,使阴性和阳性血浆样本的区分变得复杂。在目前的研究中,我们开发了一种酸变性方法,用于含有 FVIII 的血浆样本,有助于识别含有 FVIII 特异性抗体的样本,并比较了加热和酸变性对单克隆环境下检测 FVIII-抗体相互作用的影响。我们研究的目的是建立一种分析方法,以便在对 FVIII 耐受的情况下做出更安全的治疗决定。
Acid Treatment of FVIII-Containing Plasma Samples Unmasks a Broad Spectrum of FVIII-Specific Antibodies in ELISA.
During routine treatment, plasma samples of patients with hemophilia A or acquired hemophilia A are frequently analyzed for the presence of FVIII-specific antibodies. While only inhibitory antibodies can be detected by the Bethesda assay, inhibitory and non-inhibitory antibodies can be detected by ELISA. However, plasma samples of patients frequently contain endogenous or substituted FVIII, hence interfering with both types of analyses. One option for the inactivation of FVIII is heat denaturation, which unfortunately has been shown to lead to high background signals complicating the discrimination of negative and positive plasma samples. In the current study, we developed a method of acid denaturation for FVIII-containing plasma samples that can help identify samples containing FVIII-specific antibodies and compared the effects of heat and acid denaturation on the detection of FVIII-antibody interactions in a monoclonal setting. The aim of our study was to establish an analysis that allows safer treatment decisions in the context of tolerance to FVIII.
期刊介绍:
Hämostaseologie is an interdisciplinary specialist journal on the complex topics of haemorrhages and thromboembolism and is aimed not only at haematologists, but also at a wide range of specialists from clinic and practice. The readership consequently includes both specialists for internal medicine as well as for surgical diseases.