Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, 66047, USA.
Biopharmaceutical Innovation and Optimization Center, University of Kansas, Lawrence, KS, 66047, USA.
Anal Methods. 2022 Jan 20;14(3):241-249. doi: 10.1039/d1ay01832a.
This study aimed to investigate the diffusivities of lysozyme (LYS), ovalbumin (OVA), and hyaluronic acid (HA) in buffered solvents using dynamic light scattering (DLS). For protein/solvent and HA/solvent binary systems, the diffusion coefficients of protein or HA were obtained from autocorrelation function (ACF) curve fitting. Whereas, for protein/HA/solvent ternary systems, the two eigenvalues of the mutual diffusion coefficient matrix were obtained from ACF curve fitting. The results of binary systems showed that at low ionic strength, the diffusion coefficients of protein and HA increased linearly with concentration; at high ionic strength, the diffusion coefficients of OVA and LYS were independent on protein concentration; for HA, the positive linear relationship between diffusion coefficient and concentration existed at high and low ionic strengths, but the slope at high ionic strength was smaller compared to that at low ionic strength. For OVA/HA/solvent ternary systems, the sum of two eigenvalues (1DLS + 2DLS) was slightly smaller compared to ( + ), where and were the diffusion coefficients in their binary systems. On the contrary, for LYS/HA/solvent ternary systems, (1DLS + 2DLS) were significantly smaller than ( + ) and the diffusion coefficients in binary and ternary systems exhibited an opposite trend with respect to ionic strength change. The DLS and MD simulation results indicated strong attractive intermolecular interaction existed between LYS and HA molecules, especially at low ionic strength. By using DLS, it was possible to characterize the diffusion coefficients of diluted protein/HA binary and ternary systems.
本研究旨在使用动态光散射（DLS）研究溶菌酶（LYS）、卵清蛋白（OVA）和透明质酸（HA）在缓冲溶剂中的扩散系数。对于蛋白质/溶剂和 HA/溶剂二元体系，通过自相关函数（ACF）曲线拟合获得蛋白质或 HA 的扩散系数。而对于蛋白质/HA/溶剂三元体系，则通过 ACF 曲线拟合获得互扩散系数矩阵的两个特征值。二元体系的结果表明，在低离子强度下，蛋白质和 HA 的扩散系数随浓度呈线性增加；在高离子强度下，OVA 和 LYS 的扩散系数与蛋白质浓度无关；对于 HA，在高离子强度和低离子强度下，扩散系数与浓度呈正线性关系，但高离子强度下的斜率小于低离子强度下的斜率。对于 OVA/HA/溶剂三元体系，两个特征值（1DLS + 2DLS）之和略小于（ + ），其中 和 分别是其二元体系中的扩散系数。相反，对于 LYS/HA/溶剂三元体系，（1DLS + 2DLS）明显小于（ + ），并且二元和三元体系中的扩散系数随离子强度的变化呈现出相反的趋势。DLS 和 MD 模拟结果表明，LYS 和 HA 分子之间存在强烈的吸引力相互作用，特别是在低离子强度下。通过使用 DLS，可以对稀释的蛋白质/HA 二元和三元体系的扩散系数进行表征。
