本文選題：親和層析　切入點：親和介質(zhì)　出處：《北京化工大學(xué)》2009年碩士論文　論文類型：學(xué)位論文

【摘要】： 論文對乙肝人免疫球蛋白(HHBIG)和狂犬人免疫球蛋白(HRIG)兩種特異性免疫球蛋白的分離純化方法進行了研究,針對親和3號配基這樣的生物大分子物質(zhì)制備親和介質(zhì)所遇到的配基密度低的難題進行了探索。主要工作如下: (1)通過對HHBIG原料進行物性分析,確定采用親和層析方法進行HHBIG的純化。以瓊脂糖凝膠為基質(zhì)制備了用于純化乙肝免疫球蛋白的親和層析介質(zhì)Sepharose 4FF-HHBIG,并優(yōu)化了HHBIG親和層析的操作條件。對效價為95 IU/mL的HHBIG原料,純化倍數(shù)達到30左右;活性收率達到45%以上;對效價為45 IU/mL的HHBIG原料,純化倍數(shù)達到40左右,活性收率達到50%以上。通過在HHBIG親和層析單元操作中加入2%PEG D作為純化伴侶保護HHBIG的活性,使得HHBIG的活性回收率提高10-15%。 (2)以瓊脂糖凝膠為基質(zhì)制備了用于純化狂犬免疫球蛋白的親和層析介質(zhì)Sepharose 4FF-HRIG,并對親和層析條件進行了優(yōu)化,對狂犬免疫球蛋白的純化倍數(shù)在15左右,活性收率達到18%。 (3)以改性PS大孔微球制備了用于純化狂犬免疫球蛋白的新型親和介質(zhì)mPS-HRIG,并將mPS-HRIG和Sepharose 4FF-HRIG親和介質(zhì)的理化性能和層析效果進行了比較。新型mPS-HRIG親和介質(zhì)的配基密度約為Sepharose 4FF-HRIG親和介質(zhì)的兩倍,mPS-HRIG親和介質(zhì)分離純化的活性回收率和純化倍數(shù)高于Sepharose 4FF-HRIG親和介質(zhì),且mPS-HRIG親和介質(zhì)的機械強度高,是一種理想的分離純化HRIG的親和層析介質(zhì)。
[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.
【學(xué)位授予單位】：北京化工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號】：R392.1

【引證文獻】

相關(guān)博士學(xué)位論文 前1條

1 湯潔莉;蛋膜光化學(xué)傳感器的研究[D];吉林大學(xué);2011年

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本文編號：1630068