地高辛标记探针检测重组RMBAYL多肽DNA残留量的研究

Study of Residual DNA in Recombinant RMBAYL by Using Digoxin Labeled Probe

为了建立标记探针检测基因重组RMBAYL多肽DNA残留量的方法,以基因重组技术和固相柱纯化技术制备重组PACAP衍生多肽RMBAYL,以工程菌总DNA为模板,用地高辛标记探针,以此探针杂交并进行显色反应。实验结果表明,3批重组RMBAYL半成品中每人份剂量的外源性DNA残留量均小于100 Pg。研究的实验方案可实现重组PACAP衍生多肽RMBAYL的高效表达和纯化,地高辛标记探针技术检测外源性DNA灵敏度高、稳定性强,可用于RMBAYL等重组PACAP衍生多肽制备过程中的质量监控及半成品的快速、高效检定。

To determine the residual DNA in recombinant RMBAYL by using digoxin labeled probe and establish a method for analyzing the residual DNA in recombinant PACAP derived peptide, recombinant RMBAYL was prepared by genetic engineering technique and solid-phase purification technique. Total DNA of engineering bacteria of recombinant RMBAYL was labeled by digoxin and used as probe for hybridization assay of residual DNA in semi-finished products of recombinant RMBAYL for injection. The experiment results showed that the residual DNA in three batches of semi-finished products was less than 100pg per dosage for one person. The current methods not only could achieve highly efficient expression and purification of recombinant RMBAYL, but also digoxin labeled probe method for determining residual DNA in recombinant RMBAYL had high sensitivity and stability and it could be used for the quality control of recombinant PACAP derived peptide or specific, rapid and efficient detection of semi-finished products.