特异性人免疫球蛋白分离纯化方法研究

发布时间：2018-03-18 14:55

本文选题：亲和层析　切入点：亲和介质　出处：《北京化工大学》2009年硕士论文　论文类型：学位论文

【摘要】： 论文对乙肝人免疫球蛋白(HHBIG)和狂犬人免疫球蛋白(HRIG)两种特异性免疫球蛋白的分离纯化方法进行了研究,针对亲和3号配基这样的生物大分子物质制备亲和介质所遇到的配基密度低的难题进行了探索。主要工作如下: (1)通过对HHBIG原料进行物性分析,确定采用亲和层析方法进行HHBIG的纯化。以琼脂糖凝胶为基质制备了用于纯化乙肝免疫球蛋白的亲和层析介质Sepharose 4FF-HHBIG,并优化了HHBIG亲和层析的操作条件。对效价为95 IU/mL的HHBIG原料,纯化倍数达到30左右;活性收率达到45%以上;对效价为45 IU/mL的HHBIG原料,纯化倍数达到40左右,活性收率达到50%以上。通过在HHBIG亲和层析单元操作中加入2%PEG D作为纯化伴侣保护HHBIG的活性,使得HHBIG的活性回收率提高10-15%。 (2)以琼脂糖凝胶为基质制备了用于纯化狂犬免疫球蛋白的亲和层析介质Sepharose 4FF-HRIG,并对亲和层析条件进行了优化,对狂犬免疫球蛋白的纯化倍数在15左右,活性收率达到18%。 (3)以改性PS大孔微球制备了用于纯化狂犬免疫球蛋白的新型亲和介质mPS-HRIG,并将mPS-HRIG和Sepharose 4FF-HRIG亲和介质的理化性能和层析效果进行了比较。新型mPS-HRIG亲和介质的配基密度约为Sepharose 4FF-HRIG亲和介质的两倍,mPS-HRIG亲和介质分离纯化的活性回收率和纯化倍数高于Sepharose 4FF-HRIG亲和介质,且mPS-HRIG亲和介质的机械强度高,是一种理想的分离纯化HRIG的亲和层析介质。
[Abstract]:In this paper, the isolation and purification of two specific immunoglobulin (HHBG) and rabies human immunoglobulin (HRIGG) were studied. The problem of low ligand density in the preparation of affinity medium for biomolecules such as affinity ligand 3 was explored. The main work is as follows:. 1) by analyzing the physical properties of HHBIG raw materials, HHBIG was purified by affinity chromatography. Sepharose _ 4FF-HHBIGs, an affinity chromatography medium used to purify hepatitis B immunoglobulin, was prepared by agarose gel, and the operating conditions of HHBIG affinity chromatography were optimized. The purification multiple is about 30, the activity yield is over 45%, and the purification multiple is about 40 for the HHBIG with a titer of 45 IU/mL. The activity yield was over 50%. By adding 2PEGD into the HHBIG affinity chromatography unit to protect the activity of HHBIG, the recovery rate of HHBIG was increased 10-15%. (2) the affinity chromatography medium Sepharose 4FF-HRIGI was prepared by agarose gel as the matrix for purification of rabies immunoglobulin, and the affinity chromatography conditions were optimized. The purification multiple of rabies immunoglobulin was about 15, and the activity yield was 18%. A novel affinity medium mPS-HRIGI for purification of rabies immunoglobulin was prepared by modified PS macroporous microspheres. The physicochemical properties and chromatographic effects of mPS-HRIG and Sepharose 4FF-HRIG affinity media were compared. The ligand density of the new mPS-HRIG affinity medium was approximately. The activity recovery and purification multiple of Sepharose 4FF-HRIG affinity medium was higher than that of Sepharose 4FF-HRIG affinity medium. The affinity medium of mPS-HRIG has high mechanical strength and is an ideal affinity chromatography medium for the separation and purification of HRIG.
【学位授予单位】：北京化工大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392.1

【引证文献】