A purification system was constructed with the N-segment of the Npu DnaE split intein as an affinity ligand immobilized onto an epoxy-activated medium and the C-segment used as the cleavable tag fusing target protein. The affinity properties of C-tagged proteins adsorbed on I_N affinity chromatography medium were studied with GFP as a model target protein. The saturated adsorption capacity and dynamic adsorption capacity reached 51.9–21.0 mg mL⁻¹, respectively. With this system, two model proteins, GFP and alcohol dehydrogenase (ADH), has been successfully taglessly purified with regulation of Zn²⁺ and DTT. The yield, purification factor and purity of purified tagless GFP reached 39, 11.7 and 97%, respectively; while these values for purified tagless ADH were 38.2, 6.8 and 91%, respectively. These results showed that the system for Npu DnaE split intein-mediated affinity adsorption and in situ cleavage is a potential platform for recombinant protein production.

构建纯化系统，其中Npu DnaE 分裂内含肽的 N 段作为亲和配体固定在环氧活化介质上，C 段用作可切割的标签融合靶蛋白。用 GFP 作为模型靶蛋白研究了吸附在 I _N亲和层析介质上的 C 标记蛋白的亲和特性。饱和吸附容量和动态吸附容量分别达到51.9-21.0 mg mL ^-1。使用该系统，两种模型蛋白，GFP 和乙醇脱氢酶 (ADH)，已成功通过调节 Zn ^{2+ 进行无}标记纯化和 DTT。纯化的无标签GFP的产率、纯化因子和纯度分别达到39%、11.7%和97%；而纯化的无标签 ADH 的这些值分别为 38.2、6.8 和 91%。这些结果表明，Npu DnaE 分裂内含肽介导的亲和吸附和原位裂解系统是重组蛋白生产的潜在平台。