Imaging of peptide adsorption to microfluidic channels in a plastic compact disc using a positron emitting radionuclide
Imaging of peptide adsorption to microfluidic channels in a plastic compact disc using a positron emitting radionuclide
复制标题
DOI：
10.1039/b418715f
复制
发表时间：
2005-01-01
期刊：
影响因子：
6.1
通讯作者：
Wallenborg, S
中科院分区：
文献类型：
--
作者：
Lavén, M;Velikyan, I;Wallenborg, S
描述了一种通过放射性核素成像研究微流体通道内肽-表面相互作用的方法。由于微生物装置中通道的表面积与体积比高，加上分析物含量低，非特异性肽吸附是一个关键问题。因此，本研究的目的是开发一种能够直接检测微流体通道内吸附的肽的方法。选择由塑料光盘中模制的通道组成的微型装置用于研究，以及两种不同长度和等电点（pI）值的选定肽。一个双功能螯合剂，DOTA，连接到肽的共轭和短寿命的正电子发射放射性核素Ga-68标记。使用PhosphorImager系统获得微流体通道内放射性示踪剂分布的定量图像。当改变通常影响肽吸附的许多参数时，该方法的功效通过清楚地测量吸附变化的能力来证明。这些包括表面修饰、分析物浓度、pH和离子强度。此外，两种定量方法的开发和比较。放射性核素成像也允许可视化的吸附和释放过程中的微通道色谱柱。结果表明，放射性核素成像是一种合适的工具，不仅用于研究肽吸附到本研究中提出的微通道，但也作为一种通用的工具，以测量肽-表面相互作用在各种各样的固定结构和设备。
A method for studying peptide - surface interactions within microfluidic channels by radionuclide imaging is described. With the high surface area-to-volume ratio of channels in miniaturised devices, combined with low amounts of analyte, non-specific peptide adsorption is a critical issue. The objective of the study was therefore to develop a method capable of direct detection of adsorbed peptide within microfluidic channels. A micro-device consisting of channels moulded in a plastic compact disc was chosen for the study, together with two selected peptides of different lengths and isoelectric point (pI) values. A bifunctional chelator, DOTA, was attached to the peptide by conjugation and labelled with the short-lived positron emitting radionuclide Ga-68. Quantitative images of radiotracer distribution within the microfluidic channels were obtained using a PhosphorImager system. The power of the method was demonstrated by the ability to clearly measure changes in adsorption when varying a number of parameters that typically affect peptide adsorption. These included surface modifications, analyte concentration, pH, and ionic strength. Additionally, two quantification methods were developed and compared. Radionuclide imaging also permitted visualisation of adsorption and release processes in microchannel chromatographic columns. The results suggest that radionuclide imaging is a suitable tool not only for the study of peptide adsorption to the microchannels presented in this study but also as a versatile tool to measure peptide - surface interactions in a wide variety of miniaturised structures and devices.