摘要
目的研制血浆或细胞培养上清可溶性白细胞介素６受体糖蛋白１３０（ｓｇｐ１３０）的酶联免疫吸附测定（ＥＬＩＳＡ）检测国产试剂盒，为ｓｇｐ１３０ＥＬＩＳＡ的检测提供便利条件。方法单克隆抗体（单抗）经生物素标记后，采用双抗体夹心法，显色系统选用辣根过氧化物酶（ＨＲＰ）亲和素／邻苯二胺（ＯＰＤ）溶液，对７株小鼠抗人ｓｇｐ１３０单抗进行不同的组合配对，选择最佳配对用以检测ｓｇｐ１３０。结果以单抗ＢＴ１２包被，二抗为生物素标记的ＢＴ２。这一配对可用于血浆或细胞培养上清ｓｇｐ１３０水平的检测，检测灵敏度为３．１２ｎｇ／ｍｌ。结论利用识别ｓｇｐ１３０不同表位的２株单抗可建立其ＥＬＩＳＡ检测方法。本法灵敏度高，方法简便，适合临床使用。
Objective To establish an ELISA method for detecting levels of soluble gp130 (sgp130) in plasma (or cell culture supernatant) and create a sgp130 ELISA kit.Methods Labeled with biotin, the best couple of MAbs detecting sgp130 can be chosen among 7 MAbs (anti sgp130) using sandwich method. The optical system is HRP biotin and OPD solution.Results BT12 (MAb for coating) and biotin labeled BT2 *(the second MAb) were the best couple to detect sgp130 levels in plasma (or cell culture supernatant). The sensitivity of this method were about 3.12 ng/ml.Conclusion Using 2 MAbs which recognize the deferent epitopes of sgp130, we can establish he ELISA method for detecting sgp130. Because of its high sensitivity and easy operation, this method can be used in clinical laboratory.
基金
国家自然科学基金
关键词
试剂盒
受体
白细胞介素6
β糖蛋白链
ELISA
Membrane glycoproteins Receptors, interleukin Antibodies, monoclonal Reagent kits, diagnostic
