SEPARATION OF IMMUNOGLOBULIN AND TRANSFERRIN FROM BLOOD-SERUM AND PLASMA BY METAL CHELATE INTERACTION CHROMATOGRAPHY
SEPARATION OF IMMUNOGLOBULIN AND TRANSFERRIN FROM BLOOD-SERUM AND PLASMA BY METAL CHELATE INTERACTION CHROMATOGRAPHY
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DOI：
10.3168/jds.s0022-0302(88)79742-8
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发表时间：
1988-07-01
影响因子：
3.5
通讯作者：
NAKAI, S
中科院分区：
文献类型：
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作者：
ALMASHIKHI, SA;NAKAI, S
采用金属螯合相互作用色谱法分离血清和血浆中的Ig、转铁蛋白和白蛋白。柱内填充亚氨基二乙酸：1,4-丁二醇二缩水甘油酯Sepharose 6B或Sephacryl S-300，并装入铜、锌、镍或钴离子。径向免疫扩散试验表明，Zn-、Ni-、Co-和cu -负载柱的富IgG血清分别含有23.2、81.3、79.4和98.1%的活性IgG。转铁蛋白从第二峰中恢复。当同样条件的金属螯合相互作用层析用于血浆时，血红蛋白倾向于与cu负载柱强烈结合，并且仅用50%乙醇洗脱。用焦碳酸二乙酯修饰Ig和转铁蛋白中的组氨酸残基几乎完全破坏了它们与色谱柱的结合能力。分离得到的免疫球蛋白G具有抗大肠杆菌、鼠伤寒沙门菌和副百日咳博德氏杆菌的抗脂多糖抗体活性。
Metal chelate interaction chromatography was used to separate Ig, transferrin, and albumin from blood serum and blood plasma. A column was packed with iminodiacetic acid: 1,4-butanediol diglycidyl Sepharose 6B or Sephacryl S-300 and loaded with copper, zinc, nickel, or cobalt ion. Radial immunodiffusion assay indicated that Ig-rich fractions of blood serum obtained from Zn-, Ni-, Co-, and Cu-loaded columns contained 23.2, 81.3, 79.4 and 98.1% active IgG, respectively. Transferrin was recovered from the second peak. When the same conditions of metal chelate interaction chromatography were used for blood plasma, hemoglobin tended to bind strongly to the Cu-loaded column and was eluted only with 50% ethanol. Modification of histidine residues in Ig and transferrin with diethyl pyrocarbonate almost completely destroyed their binding ability to the column. Immunoglobulin G separated showed antilipopolysaccharide antibody activity against Escherichia coli, Salmonella typhimurium, and Bordettella parapertussis.