Bio Process Research and Development Laboratories, Production Division, Kyowa Hakko Kirin, Co., Ltd., Gunma, Japan.
J Sep Sci. 2010 Jan;33(1):37-45. doi: 10.1002/jssc.200900543.
The aim of this study was to classify the retention time of recombinant human monoclonal antibodies (rhMAbs) in hydroxyapatite chromatography with sodium-phosphate gradient elution according to their physicochemical properties. We analyzed 37 rhMAbs and found that (i) the structures of both the constant and variable regions affected retention time, (ii) the number of basic amino acid residues in the variable region, particularly in the heavy chain, correlated well with retention time, and (iii) this correlation was more pronounced (e.g. r(2)=0.87 for 18 kappa IgG(1) rhMAbs) when the surface accessibility of those residues are taken into account. These findings provide a useful guide for investigators and purification-process developers working with monoclonal antibodies.
本研究旨在根据重组人单克隆抗体（rhMAb）的理化性质，对羟基磷灰石层析中的保留时间进行分类，采用磷酸盐梯度洗脱。我们分析了 37 种 rhMAb，发现：（i）恒定区和可变区的结构均影响保留时间；（ii）可变区（尤其是重链）中碱性氨基酸残基的数量与保留时间密切相关；（iii）当考虑这些残基的表面可及性时，相关性更为显著（例如，18kDa κ IgG(1) rhMAb 的 r(2)=0.87）。这些发现为从事单克隆抗体研究和纯化工艺开发的研究人员提供了有用的指导。
