Mobile Affinity Sorbent Chromatography
Mobile Affinity Sorbent Chromatography
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DOI：
10.1021/acs.analchem.7b03117
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发表时间：
2018-02-06
影响因子：
7.4
通讯作者：
Regnier, Fred E.
中科院分区：
文献类型：
--
作者：
Li, ZhiYu;Kim, JinHee;Regnier, Fred E.
常规分析的目的通常是测定少量分析物。对于含有类似于10（3）个或更多组分的样品，将没有足够的峰容量将分析物与非分析物分离。本文通过一种新型的分离机制解决了这个问题，其中小的目标分析物基团以高亲和力结合到由类似于25 nm斯托克斯半径的亲水聚合物核（HPC）组成的可溶性分析物螯合传输相（ASTP）。当引入到30 nm孔径尺寸排阻色谱（SEC）柱中时，ASTP/分析物复合物在几分钟内聚集在一起，未保留，并且在第一个色谱峰中相对纯。相反，非分析物进入填充材料的孔基质，在洗脱速度上被延迟，并且稍后被洗脱，与分析物分离。通过将抗体或一些其它亲和选择剂共价偶联到高分子量HPC来实现ASTP的制造。除了螯合分析物之外，ASTP的功能是充当分子量转移剂，将比非分析物大得多的有效分子量传递给分析物，并使其在SEC空隙体积中聚集。这种分离模式被称为移动的亲和吸附层析（MASC）。纯化后，通过荧光光谱法检测ASTP/分析物复合物。
The objective in routine analyses is generally to determine a small number of analytes. With samples containing similar to 10(3) or more components there will be insufficient peak capacity to resolve analytes from nonanalytes. This issue was addressed herein through a new type of separation mechanism in which small groups of targeted analytes are bound with high affinity to a soluble analyte-sequestering transport phase (ASTP) composed of a similar to 25 nm Stokes radius hydrophilic polymer core (HPC). When introduced into a 30 nm pore diameter size-exclusion chromatography (SEC) column, ASTP/analyte complexes elute within minutes, together, unretained, and relatively pure in the first chromatographic peak. Nonanalytes, in contrast, enter pore matrices of the packing material, are retarded in elution velocity, and are eluted later, separated from analytes. Fabrication of ASTPs was achieved by covalently coupling an antibody or some other affinity selector to a high molecular weight HPC. Beyond sequestering analytes, the function of ASTPs is to act as a molecular weight shifting agent, conveying an effective molecular weight to analytes that is much larger than that of nonanalytes and causing them to elute in the SEC void volume. This mode of separation is referred to as mobile affinity sorbent chromatography (MASC). Subsequent to their purification, ASTP/analyte complexes were detected by fluorescence spectrometry.