CN106186083A - 一种具有间隔臂的微米级超顺磁磁珠的制备方法 - Google Patents
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Abstract
本发明公开了一种具有间隔臂的微米级超顺磁磁珠的制备方法,具体步骤如下:步骤一,超顺磁Fe3O4微米磁珠的制备;步骤二,磁珠表面活化;步骤三,偶联间隔臂分子。该制备工艺在纳米微球的表面引入了间隔臂,降低了偶联大分子的空间位阻,提高了大分子的活性和利用率,减少了大分子和磁珠表面的非特异性相互作用,同时该磁珠的粒径通过配比设定可以实现在一定范围内的可控制备(2‑5微米),制备工艺所用仪器设备简单经济,粒径可调且分散性好,不易聚集;表面羟基密度可以调节,进而可以调节间隔臂的长度和表面密度;间隔臂末端为羧基基团,可以与含有氨基、羟基等分子发生反应,克服了磁珠反应活性低的缺点,扩大了此微米磁珠的范围,使用效果好。
Description
技术领域
本发明涉及一种磁珠的制备方法,具体是一种具有间隔臂的微米级超顺磁磁珠的制备方法。
背景技术
微米级磁珠由于具有超顺磁特性,只有在外加磁场作用下显示条件性磁性,而且形状规则,尺寸小、比表面积大和表面可以进行各种修饰改性等特点,在生物试剂和医疗用品等领域具有广泛的应用,特别在高通量生物科学领域等技术前沿地带具有明显的技术优势。但是蛋白质等生物大分子由于具有很大的分子量,会附加一定的空间结构,因此都具有较大的分子体积。在以磁珠为载体的表面偶联生物大分子时,其空间结构会造成一定的空间位阻,从而降低磁珠表面生物大分子的加载量和生物活性,目前市场中存在的微米级磁珠都存在加载量不足的问题。
发明内容
本发明的目的在于提供一种具有间隔臂的微米级超顺磁磁珠的制备方法,以解决上述背景技术中提出的问题。
为实现上述目的,本发明提供如下技术方案:
一种具有间隔臂的微米级超顺磁磁珠的制备方法,具体步骤如下:
步骤一,超顺磁Fe3O4微米磁珠的制备:采用共沉淀法合成超顺磁Fe3O4微米磁珠:将FeCl3·6H2O和FeSO4·7H2O分别溶于HCl溶液中,浓度均为1mol/L,将两种溶液进行机械搅拌混合,通入氮气保护,加入氨水,使溶液pH值达到11,在室温下剧烈搅拌20-30分钟,以钕铁硼磁铁进行磁分离,分别用高纯水和无水乙醇依次清洗各2-5次,50-60℃真空干燥10-14小时;
步骤二,磁珠表面活化:将步骤一中的微米磁珠用电位滴定法测定微球羟基密度,分别加入与测得的羟基密度等量的1-(3-二甲基氨基丙基)-3-乙基二亚胺盐酸盐和N-羟基硫代琥珀酰亚胺,室温反应20-60分钟,将磁珠表面的羟基活化;
步骤三,偶联间隔臂分子:向步骤二中制得的磁珠中加入间隔臂分子,其加入量为磁珠羟基密度的1-3倍,在35-40℃反应3-12小时;反应结束后,对水透析除去剩余的底物,收集此微米磁珠,于0-4℃保存。
作为本发明进一步的方案:间隔臂分子中含有碳原子的数量为8个,间隔臂分子为带有氨基和羧基的化合物。
与现有技术相比,本发明的有益效果是:该制备工艺在纳米微球的表面引入了间隔臂,降低了偶联大分子的空间位阻,提高了大分子的活性和利用率,减少了大分子和磁珠表面的非特异性相互作用,同时该磁珠的粒径通过配比设定可以实现在一定范围内的可控制备(2-5微米),制备工艺所用仪器设备简单经济,粒径可调且分散性好,不易聚集;表面羟基密度可以调节,进而可以调节间隔臂的长度和表面密度;间隔臂末端为羧基基团,可以与含有氨基、羟基等分子发生反应,克服了磁珠反应活性低的缺点,扩大了此微米磁珠的范围,使用效果好。
附图说明
图1为具有间隔臂的微米级超顺磁磁珠的制备方法中制备的微米磁珠的结构式图。
图2为具有间隔臂的微米级超顺磁磁珠的制备方法中平均粒径为2.8微米的磁珠的粒度分布图。
其中:M为微米级超顺磁磁珠,n=1-3。
具体实施方式
下面结合具体实施方式对本专利的技术方案作进一步详细地说明。
实施例1
一种具有间隔臂的微米级超顺磁磁珠的制备方法,具体步骤如下:
步骤一,超顺磁Fe3O4微米磁珠的制备:采用共沉淀法合成超顺磁Fe3O4微米磁珠:将FeCl3·6H2O和FeSO4·7H2O分别溶于HCl溶液中,浓度均为1mol/L,将两种溶液进行机械搅拌混合,通入氮气保护,加入氨水,使溶液pH值达到11,在室温下剧烈搅拌20分钟,以钕铁硼磁铁进行磁分离,分别用高纯水和无水乙醇依次清洗各3次,50℃真空干燥14小时;
步骤二,磁珠表面活化:将步骤一中的微米磁珠用电位滴定法测定微球羟基密度,分别加入与测得的羟基密度等量的1-(3-二甲基氨基丙基)-3-乙基二亚胺盐酸盐和N-羟基硫代琥珀酰亚胺,室温反应20分钟,将磁珠表面的羟基活化;
步骤三,偶联间隔臂分子:向步骤二中制得的磁珠中加入间隔臂分子,间隔臂分子中含有碳原子的数量为8个,间隔臂分子为带有氨基和羧基的化合物,其加入量为磁珠羟基密度的2倍,在35℃反应5小时;反应结束后,对水透析除去剩余的底物,收集此微米磁珠,于2℃保存。
实施例2
一种具有间隔臂的微米级超顺磁磁珠的制备方法,具体步骤如下:
步骤一,超顺磁Fe3O4微米磁珠的制备:采用共沉淀法合成超顺磁Fe3O4微米磁珠:将FeCl3·6H2O和FeSO4·7H2O分别溶于HCl溶液中,浓度均为1mol/L,将两种溶液进行机械搅拌混合,通入氮气保护,加入氨水,使溶液pH值达到11,在室温下剧烈搅拌25分钟,以钕铁硼磁铁进行磁分离,分别用高纯水和无水乙醇依次清洗各4次,55℃真空干燥10小时;
步骤二,磁珠表面活化:将步骤一中的微米磁珠用电位滴定法测定微球羟基密度,分别加入与测得的羟基密度等量的1-(3-二甲基氨基丙基)-3-乙基二亚胺盐酸盐和N-羟基硫代琥珀酰亚胺,室温反应40分钟,将磁珠表面的羟基活化;
步骤三,偶联间隔臂分子:向步骤二中制得的磁珠中加入间隔臂分子,间隔臂分子中含有碳原子的数量为8个,间隔臂分子为带有氨基和羧基的化合物,其加入量为磁珠羟基密度的3倍,在40℃反应8小时;反应结束后,对水透析除去剩余的底物,收集此微米磁珠,于4℃保存
实施例3
一种具有间隔臂的微米级超顺磁磁珠的制备方法,具体步骤如下:
步骤一,超顺磁Fe3O4微米磁珠的制备:采用共沉淀法合成超顺磁Fe3O4微米磁珠:将FeCl3·6H2O和FeSO4·7H2O分别溶于HCl溶液中,浓度均为1mol/L,将两种溶液进行机械搅拌混合,通入氮气保护,加入氨水,使溶液pH值达到11,在室温下剧烈搅拌30分钟,以钕铁硼磁铁进行磁分离,分别用高纯水和无水乙醇依次清洗各5次, 60℃真空干燥10-14小时;
步骤二,磁珠表面活化:将步骤一中的微米磁珠用电位滴定法测定微球羟基密度,分别加入与测得的羟基密度等量的1-(3-二甲基氨基丙基)-3-乙基二亚胺盐酸盐和N-羟基硫代琥珀酰亚胺,室温反应60分钟,将磁珠表面的羟基活化;
步骤三,偶联间隔臂分子:向步骤二中制得的磁珠中加入间隔臂分子,间隔臂分子中含有碳原子的数量为8个,间隔臂分子为带有氨基和羧基的化合物,其加入量为磁珠羟基密度的1倍,在35-40℃反应10小时;反应结束后,对水透析除去剩余的底物,收集此微米磁珠,于3℃保存
实施例4
一种具有间隔臂的微米级超顺磁磁珠的制备方法,具体步骤如下:
步骤一,超顺磁Fe3O4微米磁珠的制备:采用共沉淀法合成超顺磁Fe3O4微米磁珠:将FeCl3·6H2O和FeSO4·7H2O分别溶于HCl溶液中,浓度均为1mol/L,将两种溶液进行机械搅拌混合,通入氮气保护,加入氨水,使溶液pH值达到11,在室温下剧烈搅拌20-30分钟,以钕铁硼磁铁进行磁分离,分别用高纯水和无水乙醇依次清洗各2次,60℃真空干燥14小时;
步骤二,磁珠表面活化:将步骤一中的微米磁珠用电位滴定法测定微球羟基密度,分别加入与测得的羟基密度等量的1-(3-二甲基氨基丙基)-3-乙基二亚胺盐酸盐和N-羟基硫代琥珀酰亚胺,室温反应60分钟,将磁珠表面的羟基活化;
步骤三,偶联间隔臂分子:向步骤二中制得的磁珠中加入间隔臂分子,间隔臂分子中含有碳原子的数量为8个,间隔臂分子为带有氨基和羧基的化合物,其加入量为磁珠羟基密度的3倍,在38℃反应12小时;反应结束后,对水透析除去剩余的底物,收集此微米磁珠,于4℃保存。
偶联间隔臂分子后的磁珠粒径为2-5微米。
上面对本专利的较佳实施方式作了详细说明,但是本专利并不限于上述实施方式,在本领域的普通技术人员所具备的知识范围内,还可以在不脱离本专利宗旨的前提下做出各种变化。
Claims (2)
1.一种具有间隔臂的微米级超顺磁磁珠的制备方法,其特征在于,具体步骤如下:
步骤一,超顺磁Fe3O4微米磁珠的制备:采用共沉淀法合成超顺磁Fe3O4微米磁珠:将FeCl3·6H2O和FeSO4·7H2O分别溶于HCl溶液中,浓度均为1mol/L,将两种溶液进行机械搅拌混合,通入氮气保护,加入氨水,使溶液pH值达到11,在室温下剧烈搅拌20-30分钟,以钕铁硼磁铁进行磁分离,分别用高纯水和无水乙醇依次清洗各2-5次,50-60℃真空干燥10-14小时;
步骤二,磁珠表面活化:将步骤一中的微米磁珠用电位滴定法测定微球羟基密度,分别加入与测得的羟基密度等量的1-(3-二甲基氨基丙基)-3-乙基二亚胺盐酸盐和N-羟基硫代琥珀酰亚胺,室温反应20-60分钟,将磁珠表面的羟基活化;
步骤三,偶联间隔臂分子:向步骤二中制得的磁珠中加入间隔臂分子,其加入量为磁珠羟基密度的1-3倍,在35-40℃反应3-12小时;反应结束后,对水透析除去剩余的底物,收集此微米磁珠,于0-4℃保存。
2.根据权利要求1所述的具有间隔臂的微米级超顺磁磁珠的制备方法,其特征在于,所述间隔臂分子中含有碳原子的数量为8个,间隔臂分子为带有氨基和羧基的化合物。
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