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发明公开
CN103558386A ELISA检测融合蛋白rMBP-NAP的方法 失效 - 权利终止

ELISA检测融合蛋白rMBP-NAP的方法
摘要：
本发明公开了一种ELISA检测融合蛋白rMBP-NAP的方法，包括包被、封闭、加样、加鼠MBP单抗、加酶标二抗、加入底物显色。本发明使用兔NAP多克隆抗体包被，尤其是经NAP-GST亲和层析纯化抗体，通过使用商品化的酶标二抗的夹心间接ELISA法，避免了制备酶标抗体的麻烦，增加了灵敏度，在测定时也无需经过特殊处理，简便快捷、灵敏、特异性好。另外，在本发明的工作浓度下，还可很好地对rMBP-NAP进行定量分析：批内变异系数小于10%，批间变异系数均小于15%；加入结构类似的干扰物NAP-GST后，OD值并没有升高，T检验证明没有发生显著差异；回收率也在85-115%之间。
摘要（英）：

The invention discloses a method for detecting rMBP-NAP of a fusion protein through ELISA. The method comprises the steps of coating, enclosing, sample adding, murine MBP monoclonal antibody adding, enzyme-labeled secondary antibody adding and chromophore-labeled substrate adding. A rabbit NAP polyclonal antibody is used to coat and is purified through NAP-GST affinity chromatography, and a sandwich indirect ELISA method using commercial enzyme-labeled secondary antibody avoids the trouble of the preparation of the enzyme-labeled antibody, increases the flexibility, needs no special treatment during determination, and has the characteristics of simplicity, fastness, sensitiveness and good specificity Additionally, the rMBP-NAP can be well quantitatively analyzed under a work concentration in the invention, the within-run coefficient of variability is less than 10%, and the interassay coefficient of variation is less than 15%; after an interferent NAP-GST having a similar structure is added, the OD value does not rise, and T examination proves that there is no substantial difference; and the recovery rate is in a range of 85-115%.

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