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体外大分子收集过程中的微透析采样膜性能

Microdialysis sampling membrane performance during in vitro macromolecule collection.

作者信息

Wang Xiangdan, Stenken Julie A

机构信息

Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, 4215 Biotech Building, Rensselaer Polytechnic Institute, 110 Eighth Street, Troy, NY 12180-3590, USA.

出版信息

Anal Chem. 2006 Sep 1;78(17):6026-34. doi: 10.1021/ac0602930.

Abstract

Microdialysis sampling is well-established for sampling small molecules. Recently, there has been an increased interest toward collecting macromolecules using microdialysis sampling. In this work, fluorescein isothiocyanate-labeled dextrans (FITC-dextrans) with molecular weight between 10 and 70 kDa were chosen as representative molecules to study analyte mass transport properties during microdialysis sampling using different lengths (2 and 10 mm) of 100-kDa MWCO polyethersulfone membranes. Experiments were performed in both well-stirred and quiescent phosphate-buffered saline solutions as well as in a 0.3% agar solution. Different fundamental parameters affecting microdialysis sampling of macromolecules, including effective membrane diffusion coefficients, were evaluated. The applicability of the most-often-cited Bungay et al. mass transfer model was compared to experimental data for the FITC-dextrans. For the larger macromolecules, the membrane provides a significant mass transport resistance most likely caused by hindered diffusion. These experimental aspects that are critical to microdialysis sampling of macromolecules are presented.

摘要

微透析采样在小分子采样方面已得到广泛应用。近来,人们对使用微透析采样收集大分子的兴趣日益浓厚。在这项工作中,选择分子量在10至70 kDa之间的异硫氰酸荧光素标记葡聚糖(FITC - 葡聚糖)作为代表性分子,以研究使用不同长度(2和10 mm)的100 kDa截留分子量的聚醚砜膜进行微透析采样期间分析物的质量传输特性。实验在充分搅拌的静态磷酸盐缓冲盐溶液以及0.3%琼脂溶液中进行。评估了影响大分子微透析采样的不同基本参数,包括有效膜扩散系数。将最常引用的Bungay等人的传质模型的适用性与FITC - 葡聚糖的实验数据进行了比较。对于较大的大分子,膜提供了显著的质量传输阻力,这很可能是由扩散受阻引起的。本文介绍了这些对大分子微透析采样至关重要的实验方面。

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