双位酶免疫法测定NT-3cDNA-CHO细胞上清中rNT-3含量

Determination of the levels of recombinant neurotrophin 3 in the supernatants of NT 3cDNA/CHO cells by two site enzyme immunoassay

目的：定量测定ＮＴ３ｃＤＮＡＣＨＯ 细胞培养上清中基因重组ＮＴ３ 的含量。方法：利用ＮＴ３ 的ＭｏＡｂ ，建立了双位ＥＩＡ 法。结果：该法检测限为５ ．０ｐｇｍｌ ，批内差的变异系数为６ ．２ ％ ～１１ ．４ ％ （ ｎ ＝ ６） ，批间差变异系数为５ ．４ ％ ～８ ．２ ％ （ｎ ＝ ６） 。４ 株阳性细胞培养上清中有高水平的目的产物。结论：该ＥＩＡ 法简单准确；用该法成功筛选到一最高表达基因重组ＮＴ３ 的单克隆细胞株。

OBJECTIVE:To determine the levels of recombinant neurotrophin 3(NT 3) in the supernatants of NT 3cDNA/CHO cells.METHOD:A two site enzyme immunoassay(EIA) method was established using the monoclonal antibody 3W3.RESULTS:The limit of detection of the EIA was 5.0pg NT 3/ml.The coefficients of variation for within and between assays were 6.2%～11.4%(n=6) and 5.4%～8.2%(n=6),respectively.Four strains of NT 3cDNA/CHO cells had expressed high levels of recombinant NT 3.CONCLUSION:The EIA method was simple and accurate.We successfully obtained a CHO monoclonal strain expressing the highest level of recombinant NT 3.