Augustsson Per, Persson Jonas, Ekström Simon, Ohlin Mats, Laurell Thomas
Department of Electrical Measurements, Division of Nanobiotechnology, Lund University, P.O. Box 118, S-221 00, Lund, Sweden.
Lab Chip. 2009 Mar 21;9(6):810-8. doi: 10.1039/b811027a. Epub 2008 Dec 1.
Highly efficient washing and extraction of microbeads to decomplex analytes ranging from small peptides to large viruses was realised in a microscaled continuous flow format. The bead washing principle reported herein is based on acoustophoresis, i.e. the primary acoustic radiation force in an ultrasonic standing wave and laminar flow properties are utilised to translate bioanalytes trapped on functionalised microbeads from one carrier fluid to another. The carry-over of non-specific material ranges from 1 to 50 ppm relative to input levels depending on application, making acoustophoresis suitable for extraction of rare species from complex environments. Selective extraction of a phosphopeptide relative to its unphosphorylated counterpart is demonstrated using metal oxide affinity capture (MOAC) beads and MALDI-TOF MS readout. Acoustophoresis of microbeads activated with specific binders could be used to capture phage viral particles. The efficiency of the acoustophoretic washing principle was demonstrated by an unspecific phage cross contamination level of only 10(-6) of that in the input bead/phage mixture. The continuous flow format makes acoustophoretic washing flexible regarding sample volume and also allows for easy integration into a sequence of particle handling and analytical unit operations.
在微尺度连续流动模式下实现了对微珠的高效洗涤和萃取,以解聚从小肽到大型病毒等各种分析物。本文报道的微珠洗涤原理基于声泳,即利用超声驻波中的初级声辐射力和层流特性,将捕获在功能化微珠上的生物分析物从一种载液转移到另一种载液。根据应用的不同,非特异性物质的残留量相对于输入水平在1至50 ppm之间,这使得声泳适用于从复杂环境中萃取稀有物种。使用金属氧化物亲和捕获(MOAC)微珠和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)读数,证明了相对于未磷酸化对应物对磷酸肽的选择性萃取。用特异性结合剂激活的微珠的声泳可用于捕获噬菌体病毒颗粒。声泳洗涤原理的效率通过非特异性噬菌体交叉污染水平仅为输入微珠/噬菌体混合物中的10⁻⁶得到证明。连续流动模式使声泳洗涤在样品体积方面具有灵活性,并且还便于轻松集成到一系列颗粒处理和分析单元操作中。