Influence of immobilized biomolecules on magnetic bead plug formation and retention in capillary electrophoresis.
Influence of immobilized biomolecules on magnetic bead plug formation and retention in capillary electrophoresis.
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DOI：
10.1002/elps.201100353
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发表时间：
2012-03
期刊：
影响因子：
2.9
通讯作者：
Gilman SD
中科院分区：
文献类型：
--
作者：
Henken RL;Chantiwas R;Gilman SD
研究了毛细管电泳过程中磁珠塞形成和保留的显著变化，并证明了这些影响是由固定在磁珠表面的生物分子类型引起的。通过生物素-亲和素结合，将抗体、寡核苷酸和碱性磷酸酶三种生物分子分别附着在完全相同的链霉亲和素包被的磁珠上，以分离因固定在磁珠表面的生物分子类型而导致的磁珠在磁场中的固定差异。用环氧基团将碱性磷酸酶附着在磁珠表面（代替亲和素-生物素结合），研究固定化化学的影响。利用光散射法对磁珠塞的形成和保留进行了定量研究，并用显微镜对磁珠塞的形成和保留进行了定性研究。固定在磁珠表面的生物分子类型和用于将生物分子与磁珠连接的化学物质都会影响磁珠塞的形成和保留。
Significant changes in the formation and retention of magnetic bead plugs in a capillary during electrophoresis were studied, and it was demonstrated that these effects were due to the type of biological molecule immobilized on the surface of these beads. Three biological molecules, an antibody, an oligonucleotide and alkaline phosphatase, were attached to otherwise identical streptavidin-coated magnetic beads through biotin-avidin binding in order to isolate differences in bead immobilization in a magnetic field resulting from the type of biological molecule immobilized on the bead surface. Alkaline phosphatase also was attached to the magnetic beads using epoxy groups on the bead surfaces (instead of avidin-biotin binding) to study the impact of immobilization chemistry. The formation and retention of magnetic bead plugs were studied quantitatively using light scattering detection of magnetic particles eluting from the bead plugs and qualitatively using microscopy. Both the type of biomolecule immobilized on the magnetic bead surface and the chemistry used to link the biomolecule to the magnetic bead impacted the formation and retention of the bead plugs.
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