CN1900128A - 离子交换型溶解可逆光敏聚合物 - Google Patents
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Abstract
本发明涉及一种用于生物活性物质分离或提纯的离子交换型光敏聚合物,其以下列化合物(1)、化合物(2)、化合物(3)和化合物(4)为单体经共聚而得,其中化合物(1)、化合物(2)、化合物(3)与化合物(4)的摩尔比为286∶27∶7∶1,所得共聚物的粘均分子量为103~105。本发明设计并合成的光敏感溶解可逆离子交换共聚物其在光密度大于1.70×105W/m2的激光照射的条件下(470~490nm)可再生回收(回收率达95%以上)。可用于对pH值或温度敏感的生物活性物质分离或提纯。其中:R1、R2分别选自H或C1~C6的烷基中一种;R3为C1~C6的烷基;R4为CH3或CHO;R5为COOH、COONa或COOK,R6、R7、R8分别选自H或C1~C6的烷基中一种。
Description
技术领域
本发明涉及一种溶解可逆光敏聚合物,具体地说,涉及一种用于生物活性物质分离或提纯的离子交换型光敏聚合物。
背景技术
当环境的物理场(如pH值、温度或光照等条件)发生改变,或存在特定离子、有机溶剂以及带相反电荷的聚电解质时,某些大分子聚合物的溶解状态会发生可逆性的改变,这些大分子聚合物被称为“智能”聚合物,其被广泛应用于生物化学、医药和生物化工等领域的产品分离和提纯。
发明人所在研究小组已在CN 1687162A中公开了一种用于两水相系统的光敏可再生聚合物。本发明的发明人在现有技术的基础上,设计并合成了一种用于生物活性物质分离或提纯的离子交换型光敏聚合物,丰富和拓展了溶解可逆光敏聚合物的种类及应用领域。
发明内容
本发明目的在于,提供一种离子交换型溶解可逆光敏聚合物,丰富和拓展溶解可逆光敏聚合物的种类。
本发明所说的离子交换型溶解可逆光敏聚合物,其以下列化合物(1)、化合物(2)、化合物(3)和化合物(4)为单体经共聚而得,其中化合物(1)、化合物(2)、化合物(3)与化合物(4)的摩尔比为286∶27∶7∶1,所得共聚物的粘均分子量为103~105,
其中:R1、R2分别选自H或C1~C6的烷基中一种;R3为C1~C6的烷基;R4为CH3或CHO;R5为COOH、COONa或COOK,R6、R7、R8分别选自H或C1~C6的烷基中一种。
本发明的一个优选方案是:R1、R2分别选自H或C1~C3的烷基中一种;
本发明的另一个优选方案是:R3为C1~C4的烷基;
本发明的再一个优选方案是:R6、R7、R8分别选自H或C1~C3的烷基中一种。
本发明的最佳方案为:R1为H;R2为异丙基;R3为正丁基;R6、R7、R8为H;R5为COONa;R4为CH3。
制备本发明所说共聚物的主要步骤如下:
以苯或是乙醇为溶剂,偶氮二异丁腈(AIBN)为引发剂,将化合物(1)、化合物(2)、化合物(3)和化合物(4)按摩尔比为286∶27∶7∶1加入溶剂中,振荡并用超声装置作用使化合物混合均匀,在有惰性气体存在下于50~80℃反应24小时,然后将聚合物加入正己烷至沉淀析出,所得沉淀物经过滤和干燥后即为目标物。
其中:化合物(1)是以丙烯酰氯为原料,与相应胺反应后制得;化合物(2)是由丙烯酸与相应的醇经酯化反应制得;化合物(3)与化合物(4)为市售品。
本发明设计并合成的光敏感溶解可逆离子交换共聚物其在光密度大于1.70×105W/m2的激光照射的条件下(470~490nm)可再生回收(回收率达95%以上)。可用于对pH值或温度敏感的生物活性物质分离或提纯。
具体实施方式
下面通过实施例对本发明作进一步说明,其目的仅在于更好理解本发明的内容而非限制本发明的保护范围:
实施例1
异丙基丙烯酰胺、丙烯酸丁酯、丙烯酸和叶绿酸铜钠[化合物(4)其中R5为COONa,R4为CH3]共聚物的制备:
在250ml锥形瓶中加入120ml苯液,以及引发剂AIBN(0.006mol),然后依次加入叶氯酸铜钠(0.001mol),丙烯酸(0.007mol),丙烯酸丁酯(0.027mol),NIPA(0.287mol),混合充分,摇匀。通氮15分钟以排除氧气,密封。将锥形瓶放入超声装置中作用10分钟,然后放入60℃的旋转震荡型水浴摇床中,在250rpm的转速下反应24小时。反应结束后,在瓶底形成固状聚合物,将聚合物中溶剂除去,以正己烷进行沉淀,将聚合物放入真空干燥箱内干燥至恒重,然后研磨成粉末。将聚合物溶于去离子水中,离心除去不溶固体。蒸发后真空干燥,研磨成粉末,备用。其粘均分子量为5.5×103。
实施例2
将聚合物溶解于pH值为6.2的柠檬酸缓冲液中配成浓度为6%的溶液,取1ml聚合物溶液置于光密度大于1.70×105W/m2的488nm的激光进行照射大约半小时,聚合物从溶液当中分相并沉淀,再次将聚合物溶解于pH值为6.2的柠檬酸缓冲液中,再次照射使得聚合物沉淀,循环使用聚合物八次,聚合物的回收率保持在95%~98%的范围内。按照此回收率,理论上可以使用四十次后,仍有10%的聚合物回收。
实施例3
使用pH为6.2柠檬酸-柠檬酸钠缓冲液配制浓度约为50000U/ml溶菌酶(lysozyme)溶液,用此溶菌酶溶液配制浓度为10wt%的聚合物溶液,取1ml上述溶液置入10℃且转速为100rpm的水浴摇床中反应6小时后。采用488nm激光光照导致聚合物沉淀,然后离心分离得到上清液和沉淀。将离心分离得到沉淀溶解于1ml的1M醋酸与0.025M盐酸的混合液进行解吸实验。通过对溶菌酶的吸附和解吸,溶菌酶的比活维持在11000U/mg以上,并且总活性略有提高,证明该聚合物在光照条件下对溶菌酶的比活和活性没有损害。
实施例4
将聚合物应用于从鸡蛋中提取溶菌酶,在0.03M的pH6.2柠檬酸缓冲液的条件下吸附,1M醋酸和0.025M盐酸的混合液解吸。蛋清中溶菌酶的比活从399U/mg提高到6346U/mg,活性回收率为81.3%。
Claims (5)
2、如权利要求1所述的离子交换型溶解可逆光敏聚合物,其特征在于,其中R1、R2分别选自H或C1~C3的烷基中一种。
3、如权利要求1所述的离子交换型溶解可逆光敏聚合物,其特征在于,其中R3为C1~C4的烷基。
4、如权利要求1所述的离子交换型溶解可逆光敏聚合物,其特征在于,其中R6、R7、R8分别选自H或C1~C3的烷基中一种。
5、如权利要求1~4中任意一项所述的离子交换型溶解可逆光敏聚合物,其特征在于,其中R1为H;R2为异丙基;R3为正丁基;R6、R7、R8为H;R5为COONa;R4为CH3。
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Cited By (2)
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CN101497675B (zh) * | 2009-01-06 | 2011-05-18 | 华东理工大学 | 一种pH敏感溶解可逆聚合物、其制备方法及用途 |
CN101497676B (zh) * | 2009-01-06 | 2012-07-04 | 华东理工大学 | 两种光敏感可逆溶解成相聚合物、其制备方法及形成的光敏性再生型两水相体系 |
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JP2005203281A (ja) * | 2004-01-16 | 2005-07-28 | Honda Motor Co Ltd | 燃料電池の昇温方法 |
CN1303118C (zh) * | 2005-03-25 | 2007-03-07 | 华东理工大学 | 一种用于两水相系统的光敏感可再生聚合物 |
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CN101497675B (zh) * | 2009-01-06 | 2011-05-18 | 华东理工大学 | 一种pH敏感溶解可逆聚合物、其制备方法及用途 |
CN101497676B (zh) * | 2009-01-06 | 2012-07-04 | 华东理工大学 | 两种光敏感可逆溶解成相聚合物、其制备方法及形成的光敏性再生型两水相体系 |
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