Affinity-purification of Transferrin-binding protein B under nondenaturing conditions |
| |
Authors: | Krell Tino Chevalier Michel Lissolo Ling |
| |
Institution: | Aventis Pasteur, 1541 avenue Marcel Mérieux, 69280 Marcy l'Etoile, France. tino.krell@aventis.com |
| |
Abstract: | The commonly used purification procedures for Transferrin-binding protein B (TbpB) are based on an affinity chromatography step using resins onto which human transferrin had been immobilized. These protocols involve protein elution using denaturing buffer solutions. Here we present an improved protocol which permits protein elution under nondenaturing conditions using chelating agents such as phosphate or compounds containing a pyrophosphate group. Furthermore, isothermal titration calorimetry experiments of the purified protein with holotransferrin have been shown to be a reliable method to assess the purity and activity of the purified material. |
| |
Keywords: | |
本文献已被 ScienceDirect PubMed 等数据库收录! |