Modeling and simulation of protein uptake in cation exchanger visualized by confocal laser scanning microscopy
Modeling and simulation of protein uptake in cation exchanger visualized by confocal laser scanning microscopy
复制标题
通过共焦激光扫描显微镜可视化阳离子交换器中蛋白质摄取的建模和模拟
DOI：
10.1016/j.chroma.2006.09.036
复制
发表时间：
2006-12-08
影响因子：
4.1
通讯作者：
Sun, Yan
中科院分区：
文献类型：
--
作者：
Yang, Kun;Shi, Qing-Hong;Sun, Yan
Confocal laser scanning microscopy (CLSM) has been extensively applied in the area of protein chromatography to investigate the uptake mechanism of protein in adsorbents. However, due to the light attenuation in the deeper layers of a specimen, quantitative analysis using CLSM data is still far from reality. In this work, an attenuation equation for describing the darkening of the CLSM image in the deeper scanning layers was developed. Bovine serum albumin (BSA) adsorption to SP Sepharose FF was performed by batch adsorption and micro-column chromatography on which protein concentration in single absorbents were visualized by CLSM. The parameters in the equation were estimated by fitting it to the fluorescence intensity profiles obtained at adsorption equilibrium, and then the equation was used to simulate the effect caused by the light scattering and absorption. CLSM analysis demonstrated that BSA adsorption to SP Sepharose FF followed the shrinking core pattern and was predicted reasonably well by the pore diffusion model in combination with the attenuation equation. By comparison of the CLSM data with the simulations, it shows that the attenuation equation was useful to demonstrate the validity of an intraparticle mass transport model for the estimation of intraparticle protein concentration profiles. (c) 2006 Elsevier B.V. All rights reserved.
