常温预辐射聚合法物理包埋辣根过氧化物酶的研究

STUDY ON A NEW IMMOBILIZATION METHOD OF HRP VIS PRE-IRRADIATION POLYMERIZATION AT ROOM TEMPERATURE

本文发展了一种新型的,常温下预辐射聚合物理包埋酶的方法。此法避免了酶直接受到辐照而引起的失活,且简单易行、包埋效率高,固相酶的使用性能亦很好。实验表明,常温预辐射固定化技术对辐射敏感的生物活性分子的固定化将是一种很有希望的新型物理包埋法。

It was found that the deactivation of Horse Radish Peroxidase(HRP) by gamma-ray was very severe at room temperature, but at low temperature(-78℃) it's deactivation could be controlled effectively. Radiation entrapping immobilization of HRP at low temperature was performed,but the leakage of immobilized HRP from gel made this method undesired. In this work, a new immobilization method of HRP vis pre-irradiation polymer-ization of acrylamide was developed. In this method. sample solution of acrylamideand proper ratio of crosslinking agent were being irradiated with bubbling oxygen con-tinuously and then, take the sample out of radiation field, put some unirradiated HRPinto the sample which was still not polymerized bubbling high pure nitrogen for about15 minutes. Finally the sample was polymerized and at the same time the HRP wouldbe entrapped in it. Some entrapping conditions were investigated. About 60% of HRP can be en-trapped and the leakage of HRP gel was somewhat improved.