Automated microanalysis using magnetic beads with commercial capillary electrophoretic instrumentation
Automated microanalysis using magnetic beads with commercial capillary electrophoretic instrumentation
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DOI：
10.1016/s0021-9673(97)00629-8
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发表时间：
1997-09-26
影响因子：
4.1
通讯作者：
Karger, BL
中科院分区：
文献类型：
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作者：
Rashkovetsky, LG;Lyubarskaya, YV;Karger, BL
利用磁珠（mb）和商用毛细管电泳（CE）仪器进行酶和抑制试验，以及分析生物分子（如抗原、底物等）的一种新的微量分析方法的潜力已经被探索。将少量含有固定化生物分子的磁珠注入中性亲水性涂覆的熔融二氧化硅毛细管中。短塞（2-3毫米）的珠子被放置在CE系统的墨盒中的磁铁固定，而不使用砖块。每次下入后，珠子都可以更换，无需重新生成固体支撑。分析采用两种方案：顺序注射（SI）和SI后等速电泳（ITP）聚焦。碱性磷酸酶（AP）和hiv蛋白酶被用来演示SI程序的酶和抑制试验。第二种方法是SI/ITP，使用固定在微球上的抗体（羊IgG抗体）来定量抗原（小鼠单抗）。MB-CE方法，只需要飞摩尔（fmol）量的材料，可以潜在地用于诊断和法医分析，动力学研究和寻找抑制剂，配体，受体等(C) 1997 Elsevier Science B.V.
The potential of a new microanalytical method using magnetic beads (MBs) and commercial capillary electrophoresis (CE) instrumentation for performing enzymatic and inhibition assays, as well as for analysis of biological molecules such as antigens, substrates, etc., has been explored. A small quantity of magnetic beads containing immobilized biomolecules was injected into a neutral hydrophilic-coated fused-silica capillary. The short plug (2-3 mm) of beads was held fixed by a magnet placed in the cartridge of the CE system, without the use of frits. The beads could be replaced after each run, eliminating the need to regenerate the solid support. Two protocols were used for analysis: sequential injection (SI) and SI followed by isotachophoretic (ITP) focusing. Alkaline phosphatase (AP) and HIV-protease were used to demonstrate the SI procedure for enzymatic and inhibition assays. The second protocol, SI/ITP, was employed to quantitate an antigen (mouse mAB) using antibodies (sheep IgG towards mouse AB) immobilized on the beads. The MB-CE method, requiring only femtomole (fmol) quantities of material, can potentially be employed in diagnostic and forensic assays, kinetic studies and searching for inhibitors, ligands, receptors, etc. (C) 1997 Elsevier Science B.V.