CN106955505A - 交联琼脂糖凝胶分离材料的制备 - Google Patents

交联琼脂糖凝胶分离材料的制备 Download PDF

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CN106955505A
CN106955505A CN201610011411.XA CN201610011411A CN106955505A CN 106955505 A CN106955505 A CN 106955505A CN 201610011411 A CN201610011411 A CN 201610011411A CN 106955505 A CN106955505 A CN 106955505A
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cross
preparation
gel
agarose gel
linked agarose
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赵婷
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Xi'an Jinuotai Biological Technology Co Ltd
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Xi'an Jinuotai Biological Technology Co Ltd
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
    • B01D15/34—Size selective separation, e.g. size exclusion chromatography, gel filtration, permeation
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
    • C08J2305/12—Agar-agar; Derivatives thereof

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  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Organic Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Treatment Of Liquids With Adsorbents In General (AREA)

Abstract

本发明公布了一种交联琼脂糖凝胶的制备方法，本发明所涉及的制备方法可采用多次交联的方法。本发明以二次交联为实施方案，制备一种高流速的交联琼脂糖凝胶层析介质。经过二次交联后的凝胶介质形成刚性，半刚性的产品，提高了珠体的机械稳定性和通透性，降低柱的反压，可用于生物制品的工业规模生产使用。

Description

交联琼脂糖凝胶分离材料的制备
技术领域
本发明涉及一种交联琼脂糖的分离材料，并用于制备所述分离材料的制备方法。
背景技术
凝胶过滤层析已有40多年的应用历史，在酶、蛋白质、多肽、核酸等大分子的分离纯化中起重要重用，在各种柱层析的分离中是最温和的处理方法。分离过程是基于溶解物质的扩散，而凝胶可以结合大量的水的能力。浸泡在水溶液中的不同分子尺寸大小的混合物，在不同的速率下通过凝胶床，从而达到分离纯化的目的。
凝胶过滤的生物物质分离的理想凝胶应是亲水性的，大孔，耐化学，机械稳定，不应该包含带电基团或其他吸附中心。多糖类的分子结构中含有大量的羟基，为了使凝胶更稳定，从化学和机械的角度来看，凝胶的结构必须是交联的。在凝胶分子链间的2个羟基功能团中引入含有环氧基、卤代烷基等化合物作为交联剂进行交联反应达到交叉链接时，从而使其结构稳定。
脂肪族环氧化物或相应的卤代醇或次氯代醇或卤化物作为交联剂，因为这些化合物形成醚键形成含羟基的矩阵。在色谱分离条件下，该类型的键的化学性质稳定，没有带电基团被引入的反应，达到理想的的吸附效果。
发明内容
本发明的目的在于提供一种交联琼脂糖凝胶层析分离介质的制备方法，改变琼脂糖凝胶珠体的性能，达到大规模生产要求。
本发明提供的制备方法，考虑到工业规模生产使用的需求，采用多次交联的方法，形成刚性，半刚性，大大提高了珠体的机械稳定性和通透性，降低柱的反压。
具体实施方法
(a)取6％琼脂糖凝胶珠状用去离子水洗去乙醇溶液，再依次用5倍体积20％，50％，70％，100％的二甲基亚砜混合溶液浸洗凝胶(置换出凝胶中的水)，然后将洗好的琼脂糖凝胶珠加入等体积的丙酮溶液中混合均匀，加入0.3克NaBH4，13g Na2SO4，再加入35毫升50％NaOH溶液，混合物搅拌1小时。在加入100毫升丙二醇双缩水甘油醚，继续反应过夜。反应完毕，用至少10倍体积的水洗干净。
(b)将清洗后的凝胶转移到等体积的丙酮溶液中，加入环氧氯丙烷增加反应温度到60℃和加入相当于交联剂体积的1M的氢氧化钠，逐滴加入超过6h，然后再过夜反应，反应结束后，再用大量的水进行重复清洗。
对实验所得交联琼脂糖凝胶层析分离介质做理化测试，结论层析分离介质的流速可达到350cm/h，耐压可达到0.13MPa，pH稳定性可达到3～13，符合工业规模生产使用的需求。

Claims (3)

1.一种琼脂糖凝胶的交联制备方法，其特征在于琼脂糖凝胶是采用多次交联的方法，制备成具有机械稳定性和通透性的层析介质。
2.权利要求1所述的交联琼脂糖凝胶的制备方法其特征在琼脂糖凝胶的制备是在溶剂中进行反应的，一次反应在Na2SO4，NaOH的溶液中进行反应，交联剂为长链交联剂。
3.权利要求1、2所述的交联琼脂糖凝胶的制备方法其特征在一次交联后，二次交联采用短交联剂，并其后的交联都采用短交联剂。
CN201610011411.XA 2016-01-08 2016-01-08 交联琼脂糖凝胶分离材料的制备 Pending CN106955505A (zh)

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CN106955505A true CN106955505A (zh) 2017-07-18

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Application publication date: 20170718