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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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人aβ42重组抗原的融合表达及aβ抗体的检测
, PP. 160-164
Keywords: 淀粉样β蛋白,融合表达,基因表达,克隆,分子,酶联免疫吸附测定
Abstract:
目的原核表达β-淀粉样蛋白(aβ42)抗原,建立检测aβ抗体的间接elisa技术。方法化学合成aβ42基因两个互补片段,通过pcr构建人aβ42基因,克隆至pgex-2t表达载体中,原核表达并亲和纯化gst-aβ42融合蛋白。westernblotting检测其抗原特异性。以gst-aβ42和aβ42肽分别为包被抗原,间接elisa法检测aβ42免疫的大鼠血清中aβ抗体。结果原核表达的可溶性gst-aβ42融合蛋白相对分子质量为31000,经亲和纯化后,融合蛋白产量为800mg/l菌液,纯度大于95%;westernblotting证实纯化的融合抗原与aβ单抗特异反应;以gst-aβ42为包被抗原,间接elisa法检测aβ抗体灵敏度为2ng/ml;以gst-aβ42和化学合成的aβ42分别为包被抗原,间接elisa法检测aβ42肽免疫大鼠血清中aβ抗体滴度,两者比较差异无显著性(p>0.05)。结论原核表达的gst-aβ42融合蛋白可替代昂贵的化学合成的aβ42肽抗原,用于检测aβ抗体。
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