ELECTROPHORETIC MOBILITY MODELING OF PROTEINS IN FREE ZONE CAPILLARY ELECTROPHORESIS AND ITS APPLICATION TO MONOCLONAL-ANTIBODY MICROHETEROGENEITY ANALYSIS
ELECTROPHORETIC MOBILITY MODELING OF PROTEINS IN FREE ZONE CAPILLARY ELECTROPHORESIS AND ITS APPLICATION TO MONOCLONAL-ANTIBODY MICROHETEROGENEITY ANALYSIS
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DOI：
10.1016/0021-9673(91)80085-u
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发表时间：
1991-10-18
期刊：
影响因子：
--
通讯作者：
COMPTON, BJ
中科院分区：
文献类型：
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作者：
COMPTON, BJ
推导了描述蛋白质在自由溶液中电泳迁移率的半经验模型。 如Debye-Huckel-亨利理论所述，蛋白质的迁移率被发现受到蛋白质价态、大小和形状以及溶液离子强度、pH、粘度和温度的影响。 蛋白质价，最重要的流动性决定参数固有的蛋白质，计算一个给定的pH值从其氨基酸含量使用亨德森-Hesselbalch方程。 静电荷抑制导致实际化合价低于计算值。 为了使两者相等，引入了实验确定的比例常数（F（Z））。 因此，F（Z）可以应用于蛋白质的计算价和迁移率-pH滴定曲线，从而得到蛋白质在任何给定pH下的实际迁移率。该模型进一步预测，迁移率的分子量（M）依赖性应该是M-1/3至M-2/3的连续函数，这取决于研究中的蛋白质分子量和缓冲液离子强度的大小。 该模型的许多方面都证明了其应用的免疫球蛋白G的isoelectotypes的决议，通常只解决使用等电聚焦。
A semi-empirical model for describing the electrophoretic mobility of proteins in free solution is derived. Protein mobility is found to be influenced, as dictated by the Debye-Huckel-Henry theory, by protein valence, size and shape, and by solution ionic strength, pH, viscosity and temperature. Protein valence, the most important mobility determining parameter intrinsic to the protein, is calculated for a given pH from its amino acid content using the Henderson-Hesselbalch equation. Electrostatic charge suppression causes actual valence to be less than that calculated. To equate the two an experimentally determined proportionality constant (F(Z)) is introduced. Consequently, F(Z) can be applied to the calculated valence and mobility-pH titration curve for a protein, resulting in the actual mobility of the protein at any given pH. The model further predicts that the molecular weight (M) dependency of mobility should be a continuous function of M-1/3 to M-2/3, depending on the magnitude of the protein molecular weight and buffer ionic strength under investigation. Many aspects of the model are demonstrated by its application to the resolution of immunoglobulin G isoelectrotypes, normally only resolved using isoelectric focusing.