聚乙二醇化学修饰重组人白介素-2  

Chemical Modification of Recombinant Human Interleukin-2 by Polyethylene Glycol

作　　者：王立夫[1] 吴裕忻[1] 章永平[1] 乔敏敏[1] 江石湖[1] 

机构地区：[1]上海第二医科大学附属瑞金医院消化科,200025

出　　处：《上海第二医科大学学报》1998年第1期14-18,共5页Acta Universitatis Medicinalis Secondae Shanghai

摘　　要：利用聚乙二醇活性酯（ＰＥＧ，Ｍｗ５０００）在不同反应条件下修饰重组白介素－２可改变其分子量，采用ＳＤＳ－ＰＡＧＥ测定分子量和ＣＴＬＬ－２短期培养结合＾３Ｈ－ＴｄＲ掺入法测定ＩＬ－２生物活性，ＰＥＧ修饰ｒＩＬ－２的方法学研究结果表明：随着ＰＥＧ对ｒＩＬ－２投料摩尔比的增加或修饰反应时间的延长，ｒＩＬ－２的修饰度和修饰率及修饰产物的分子量和不均一性均增加。Recombinant Human Interleukin-2 (rIL-2) is one of the cytokines with an approximate molecular weight of 15000. When rIL-2 is injected into human body, it is rapidly cleared from the circulation, resulting in short-term plasma half-life. To increase the effective molecular size, in this present investigation, we attempted to modify the rIL-2 with a water soluble and nonantigenic polymer, N-succinimidy succinate polyethylene glycol (Mw 5000), and made a comparison of different methods of chemical modifications. The extent of modification of rIL-2 by the PEG ester was determined by SDS-PAGE, and the bioactivities of IL-2 were determined by using a standard short-term proliferation response measured by [~3H] thymidine incoporated into IL-2 dependent CTLL-2 cells. Several methods for the preparation of PEG-modified rIL-2 were compared which suggested that higher molecular weight (or degree of PEG modification) and lower bioactivity of PEG-modified rIL-2 was resulted when the reaction time was longer and the concentration of PEG ester relative to rIL-2 was higher. The effective molecular size increased form 15 to 25 to 45 to 65 to 105 KDa, and the activity was inereased after renaturation of PEG-modified rIL-2. It was shown that we could apply a better coupling method for PEG-modified rIL-2 by using the optimal conditions for modification reaction.

关 键 词：RIL-2 聚乙二醇 化学修饰 免疫疗法 

分 类 号：R457.2[医药卫生—治疗学]