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Mobile Affinity Sorbent Chromatography
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-01-09 00:00:00 , DOI: 10.1021/acs.analchem.7b03117 ZhiYu Li 1 , JinHee Kim 1 , Fred E Regnier 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-01-09 00:00:00 , DOI: 10.1021/acs.analchem.7b03117 ZhiYu Li 1 , JinHee Kim 1 , Fred E Regnier 1
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The objective in routine analyses is generally to determine a small number of analytes. With samples containing ∼103 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 ∼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.
中文翻译：
移动亲和吸附色谱
常规分析的目标通常是确定少量分析物。对于含有~10 3或更多组分的样品，将没有足够的峰容量来从非分析物中分离出分析物。本文通过一种新型分离机制解决了这个问题，其中一小群目标分析物以高亲和力与由~25 nm斯托克斯半径亲水聚合物核（HPC）组成的可溶性分析物隔离运输相（ASTP）结合。当引入 30 nm 孔径尺寸排阻色谱 (SEC) 柱时，ASTP/分析物复合物在几分钟内一起洗脱，没有保留，并且在第一个色谱峰中相对纯净。相反，非分析物进入填充材料的孔基质，洗脱速度被延迟，并且稍后被洗脱，与分析物分离。 ASTP 的制造是通过将抗体或一些其他亲和选择剂与高分子量 HPC 共价偶联来实现的。除了隔离分析物之外，ASTP 的功能还可以充当分子量转移剂，将比非分析物大得多的有效分子量传递给分析物，并使它们在 SEC 空隙体积中洗脱。这种分离模式称为移动亲和吸附色谱 (MASC)。纯化后，通过荧光光谱法检测 ASTP/分析物复合物。
更新日期：2018-01-09
中文翻译：
移动亲和吸附色谱
常规分析的目标通常是确定少量分析物。对于含有~10 3或更多组分的样品，将没有足够的峰容量来从非分析物中分离出分析物。本文通过一种新型分离机制解决了这个问题，其中一小群目标分析物以高亲和力与由~25 nm斯托克斯半径亲水聚合物核（HPC）组成的可溶性分析物隔离运输相（ASTP）结合。当引入 30 nm 孔径尺寸排阻色谱 (SEC) 柱时，ASTP/分析物复合物在几分钟内一起洗脱，没有保留，并且在第一个色谱峰中相对纯净。相反，非分析物进入填充材料的孔基质，洗脱速度被延迟，并且稍后被洗脱，与分析物分离。 ASTP 的制造是通过将抗体或一些其他亲和选择剂与高分子量 HPC 共价偶联来实现的。除了隔离分析物之外，ASTP 的功能还可以充当分子量转移剂，将比非分析物大得多的有效分子量传递给分析物，并使它们在 SEC 空隙体积中洗脱。这种分离模式称为移动亲和吸附色谱 (MASC)。纯化后，通过荧光光谱法检测 ASTP/分析物复合物。
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