THE ROLE OF PROTEIN-STRUCTURE IN CHROMATOGRAPHIC BEHAVIOR
THE ROLE OF PROTEIN-STRUCTURE IN CHROMATOGRAPHIC BEHAVIOR
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DOI：
10.1126/science.3310233
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发表时间：
1987-10-16
期刊：
影响因子：
56.9
通讯作者：
REGNIER, FE
中科院分区：
文献类型：
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作者：
REGNIER, FE
色谱保留由位于多肽表面上的色谱接触区域中的相对少量的氨基酸决定。该区域由多肽内的分离模式和氨基酸分布决定。在生物亲和色谱中，接触面积可以小至几百平方埃。相比之下，在离子交换、反相、疏水相互作用和其他非生物亲和分离模式中的接触区域要宽得多，范围从多肽的一侧到整个外表面。此外，改变色谱接触区域的结构变化将改变色谱特性。因此，尽管免疫吸附剂在纯化具有相似一级结构的蛋白质方面非常有用，但它们在区分结构内的小的随机变化方面将是无效的。非生物亲和柱补充亲和柱，探测溶质表面的更大部分，并能够区分蛋白质变体。
Chromatographic retention is determined by a relatively small number of amino acids located in a chromatographic contact region on the surface of a polypeptide. This region is determined by the mode of separation and the amino acid distribution within the polypeptide. The contact area may be as small as a few hundred square angstroms in bioaffinity chromatography. In contrast, the contact region in ion exchange, reversed phase, hydrophobic interaction and the other nonbioaffinity separation modes is much broader, ranging from one side to the whole external surface of a polypeptide. Furthermore, structural changes that alter the chromatographic contact region will alter chromatographic properties. Thus, although immunosorbents can be very useful in purifying proteins of similar primary structure, they will be ineffective in discriminating between small, random variations within a structure. Nonbioaffinity columns complement affinity columns in probing a much larger portion of solute surface and being able to discriminate between protein variants.