CHARACTERIZATION OF A SOLVENT SYSTEM FOR SEPARATION OF WATER-INSOLUBLE POLIOVIRUS PROTEINS BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
CHARACTERIZATION OF A SOLVENT SYSTEM FOR SEPARATION OF WATER-INSOLUBLE POLIOVIRUS PROTEINS BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY
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DOI：
10.1016/s0021-9673(01)87434-3
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发表时间：
1985-01-01
期刊：
影响因子：
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通讯作者：
DERNICK, R
中科院分区：
文献类型：
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作者：
HEUKESHOVEN, J;DERNICK, R
高浓度的甲酸是蛋白质，特别是疏水蛋白质的一种非常有效的溶剂。在反相高效液相色谱中，60%甲酸是脊髓灰质炎病毒结构多肽和其他蛋白质增溶所必需的，在半胱氨酸处进行了修饰，与2-丙醇或乙腈一起用作梯度洗脱的有机改性剂。测试了几种反相柱。在每种情况下，定量洗脱多肽。结果表明，这种溶剂系统，其高比例的甲酸，不影响的大小，疏水性和电荷的分离多肽。通过向兔子注射脊髓灰质炎病毒多肽，在新的溶剂系统中通过色谱法获得高纯度，诱导单特异性抗体，其特异性通过免疫沉淀法测定。
Formic acid in high concentration is an extremely potent solvent for proteins, particularly for hydrophobic ones. 60% Formic acid, necessary for solubilization of structural polypeptides of poliovirus and other proteins, modified at the cysteines, was used together with 2-propanol or acetonitrile as organic modifier for gradient elution in reversed-phase high-performance liquid chromatography. Several reversed-phase columns were tested. In each case, polypeptides were eluted quantitatively. It was demonstrated that this solvent system, with its high proportion of formic acid, did not affect the size, hydrophobicity and charge of the separated polypeptides. By injection into rabbits of poliovirus polypeptides, obtained in high purity by chromatography in the new solvent system, monospecific antibodies were induced, the specificity of which was determined by immunoprecipitation.