Cytokines and Growth Factors Section, Biotherapeutics Group, National Institute for Biological Standards and Control, Health Protection Agency, Blanche Lane, South Mimms, Herts, UK.
Cytokine. 2010 May;50(2):129-37. doi: 10.1016/j.cyto.2010.01.001. Epub 2010 Jan 29.
Antibodies that neutralize interleukin-17A (IL-17A) are classically detected and quantified using cell-based assays. However, these assays are cumbersome, inherently variable and often susceptible to interference by the matrix of test samples, such as human sera. Since neutralizing antibodies block binding of IL-17A to its cell surface receptor, IL-17R, we used antibody inhibition of IL-17A binding to recombinant IL-17R/Fc fusion protein in an electrochemiluminescence (ECL) platform to develop a novel, non-cell based ligand binding assay that functionally mimics cell-based neutralization assays. Using specific polyclonal antisera and a panel of sera containing neutralizing anti-IL-17A autoantibodies from patients with autoimmune polyendocrinopathy syndrome-1, we have shown that this assay generates neutralizing titers that correlate well with those found using cell-based assays. The assay is simple to perform, reliable, and more accessible to clinical laboratories than cell-based assays. In principle, the assay methodology could be extended to detection of neutralizing antibodies to biotherapeutics for assessment of unwanted immunogenicity of biotherapeutic products.
中和白细胞介素-17A（IL-17A）的抗体通常使用基于细胞的测定法进行检测和定量。然而，这些测定法繁琐、固有变异性大，并且常常容易受到测试样品基质（如人血清）的干扰。由于中和抗体可阻断 IL-17A 与其细胞表面受体 IL-17R 的结合，因此我们使用抗体抑制 IL-17A 与重组 IL-17R/Fc 融合蛋白的结合，在电化学发光（ECL）平台上开发了一种新颖的、非基于细胞的配体结合测定法，该测定法可模拟基于细胞的中和测定法。使用特异性多克隆抗血清和一组包含自身免疫性多内分泌腺病综合征-1 患者的中和抗 IL-17A 自身抗体的血清，我们表明该测定法产生的中和效价与基于细胞的测定法一致。该测定法操作简单、可靠，并且比基于细胞的测定法更易于临床实验室使用。原则上，该测定法方法可以扩展到检测针对生物治疗药物的中和抗体，以评估生物治疗产品的不良免疫原性。
