Benedek K
AMGEN Inc., Amgen Center, Thousand Oaks, CA 91320-1789.
J Chromatogr. 1993 Aug 27;646(1):91-8. doi: 10.1016/s0021-9673(99)87010-1.
A model describing the pathway of unfolding of recombinant human brain-derived neurotrophic factor (rhBDNF) under reversed-phase high-performance liquid chromatographic conditions is introduced. The unfolding process is divided into two major steps. The first entails a series of events such as the dissociation of the dimer and the effect of the initial contact of the protein with the stationary phase. The second step is the unfolding of the monomer molecule on the hydrophobic surface exhibiting a rate constant comparable with the time scale of chromatography. The kinetics of rhBDNF unfolding is studied with respect to the type and composition of the organic solvents and as function of the temperature of chromatography. The experimental results validate the suggested multistep unfolding model. The activation energy of monomer unfolding in both solvent systems studied is similar, but the unfolding processes are different in 1-propanol and in acetonitrile.
介绍了一种描述重组人脑源性神经营养因子（rhBDNF）在反相高效液相色谱条件下展开途径的模型。展开过程分为两个主要步骤。第一步涉及一系列事件，如二聚体的解离以及蛋白质与固定相初始接触的影响。第二步是单体分子在疏水表面展开，其速率常数与色谱时间尺度相当。针对有机溶剂的类型和组成以及色谱温度，研究了rhBDNF展开的动力学。实验结果验证了所提出的多步展开模型。在所研究的两种溶剂体系中，单体展开的活化能相似，但在1-丙醇和乙腈中的展开过程不同。
