Confocal microscopy as a tool for studying protein adsorption to chromatographic matrices
Confocal microscopy as a tool for studying protein adsorption to chromatographic matrices
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DOI：
10.1016/0021-9673(96)00290-7
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发表时间：
1996-08-30
影响因子：
4.1
通讯作者：
Hjorth, R
中科院分区：
文献类型：
--
作者：
Ljunglof, A;Hjorth, R
用激光共聚焦显微镜研究了蛋白质在亲和层析基质上的吸附。通过与不同数量的荧光标记蛋白A的间歇孵育，研究了蛋白A在Ig G Sepharose6 Fast Flow上的吸附。随着样品-Ig G琼脂糖比例的增加，吸附层数也增加。同样，吸附深度依赖于孵化时间。最后，利用一组在空间上分离的共焦图像对颗粒中的吸附模式进行了三维重建。
Confocal scanning laser microscopy was used for studying protein adsorption to affinity chromatography matrices. The adsorption of Protein A to IgG Sepharose 6 Fast Flow was studied by batch incubation with varying amounts of fluorescently labeled Protein A. At low sample amounts, Protein A had been adsorbed to a thin outer layer. By increasing the sample-IgG Sepharose ratio, the adsorption layer also increased. Likewise, the adsorption depth was dependent on the incubation time. Finally, a stack of confocal images separated in space was used for three-dimensional reconstruction of the adsorption pattern in a particle.