Schriemer D C, Li L
Department of Chemistry, University of Alberta, Edmonton, Canada.
Anal Chem. 1996 Oct 1;68(19):3382-7. doi: 10.1021/ac960219p.
Molecular recognition based on the high affinity of avidin for biotin has been combined with matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The rich chemistry of biotinylation reagents underlies the development of a mass tracer method for the rapid and sensitive analysis of biotinylated analyte in complex mixtures. In this method, the biotinylated analyte is captured with immobilized avidin agarose beads. The beads with the bound complex are deposited on the MALDI sample probe, followed by elution of the retained compounds and removal of the beads. The use of MALDI matrix solutions in conjunction with drying of the beads is shown to be sufficient to overcome the avidin-biotin interaction. The studies of several different avidin-biotin interaction schemes tailored to the MALDI analysis are presented. It is demonstrated that, with an optimized scheme, the extremely high selectivity of the avidin-biotin interaction is preserved, generating species represented in the MALDI spectra that arise only from the avidin-biotin interaction. Biotinylated species in the nanomolar range can be isolated and analyzed with this technique.
基于抗生物素蛋白对生物素的高亲和力的分子识别已与基质辅助激光解吸/电离（MALDI）质谱联用。生物素化试剂丰富的化学性质为一种质量示踪方法的发展奠定了基础，该方法用于快速、灵敏地分析复杂混合物中的生物素化分析物。在这种方法中，生物素化分析物用固定化抗生物素蛋白琼脂糖珠捕获。带有结合复合物的珠子沉积在MALDI样品探头上，然后洗脱保留的化合物并去除珠子。结果表明，使用MALDI基质溶液并结合珠子干燥足以克服抗生物素蛋白 - 生物素相互作用。本文介绍了针对MALDI分析定制的几种不同抗生物素蛋白 - 生物素相互作用方案的研究。结果表明，通过优化方案，抗生物素蛋白 - 生物素相互作用的极高选择性得以保留，产生的MALDI光谱中的物种仅源于抗生物素蛋白 - 生物素相互作用。用该技术可以分离和分析纳摩尔范围内的生物素化物种。
