The monodisperse, poly(glycidylmethacrylate-co-ethylenedimethacrylate) beads with macroporous in the range of 8.0-12.0m were prepared by a single-step swelling and polymerization method. The seed particles prepared by dispersion polymerization exhibited good absorption of the monomer phase. The pore size distribution of the beads was evaluated by gel permeation chromatography and mercury instrusion method. Based on this media, a hydrophobic interaction chromatographic (HIC) stationary phase for HPLC was synthesized by a new chemically modified method. The prepared resin has advantages for biopolymer separation, high column efficiency, low column backpressure, high protein mass recovery and good resolution for proteins. The dynamic protein loading capacity of the synthesized HIC packings was 40.0mg/ml. Six proteins were fast separated in less than 8.0min using the synthesized HIC stationary phase. The HIC resin was firstly used for the purification and simultaneous renaturation of recombinant human interferon- (rhIFN-) in the extract solution containing 7.0mol/l guanidine hydrochloride with only one step. The purity and specific bioactivity of the purified of rhIFN- was found more than 95% and 1.3

本文全面地考虑了液相色谱体系中溶质和溶剂，溶质和固定相，溶荆和固定扫分子之问的各种相互怍车甸不同种类溶剂分子在固定相表面上的竞争吸咐，提出了一个具有四组参数的溶质在液相皂谱中的计量置换深留模型以文献及实验数据对比模型进行了验证结果表明，对不同的流动相和不同的固定相体系，在多组分全浓度范围内，此模型都能良好地描述实验事实。

This paper presents a retention model for proteins on an reversed-phase chro-matography support in which retention is a function of the number (Z) of solven molecules required to displace the solutte from the surface. An equation is dervied that relates the capacity factor of a protein to the displacing agent concentration and the stoichimonetry of solvent-solute displacement. Experimental tests of the model indicate that each protein has a unique Z value and that Z is directly proportional to the molecular weight of a series of protins when 60%formic acid is used as the mobile phase additive. This relationship is polypeptide solutes and the support. De-sorption curves for proteins also become more convex with incerasingly molecular weight, as predicted by the retention model. In the solvent series of methanol, etha-nol, propanol, the Z number decreases form the C1 to C3 alcohol. The Z number for any particular solvent is also related to other mobile phase additives, such as acids, and the concentration of additives.

This paper proposes a retention model which predicts that the displacement of non-polar solutes from a revesed-phase chromatographic column is accompanied by the adsortion of a stoichiometric number (Z) of solvent molecules. The number contact surface areas. Incerasing solute contact surface area would increase Z whereas increasing solvent contact surface area presented are consistent with this model. Further predictions of the model are that (1) plots of log k' versus the inverse log of solvent concentration will be non-linear at solvent concentrations where the surface of a reversed-phase support is not fully solvated, and (2) only a portion of a reversed-phase support. Non-linearity in plost of log k' versus the inverse log of solvent concentration was in fact observed at solvent concentrations where solvation of the reversed-phase support is incomplete.

With four kinds of mobile phases, methanol-water, ethanol-water, 2-propanol and acetoritrle water (all cortairing 0.1% trilfu roacetic acid), the displacemetn between soltue and solvent in RPLC wsa proved to be universal in frontal analysis (FA), Based on the measured Z value in usual RPLC to be a corstant and the quarntitative determinnation of methanol increment in mobile phase in FA. the stoichiometric displacement (SD) betwwen insulin and methanol was directly proved by the experment. The SD was also proved to ocur only on about the bonded phase layer (BPL) without any dynamic problem of mass transfer, while in FA, the SD firstly occurs on the sufaace of the BPL and then graduallly sinks into the deeper sites comparied with a dynamic problem. Although the displaced solvent by the same solute is less in the formet case, the SD in independent, of how deep of the splute enters the BPF. In addition, the numbers of the adsorbed layer on the adsorbent sur-face, but also on the exent of the complete removal of the displace-able solvent in the BPL. The physical funda mental of the SD and the methodology for investigation were also discussed.

A stoichiometric displacement retention model for proteins based on hadro-phobic interaction chromatography (HIC) is presenter. Several methods were used to demonstrate that water is the displacing agent in this process. Salt not only affects the molar concentration of water of water, but also changes the conformation of proteins, hydro-phobic interaction forces and the number of awter moiecules in a series of hydrated protein molecules. An equation that relates the capacity foactor of proteins, K', to the water concentration and the stoichiometric displacement factor, Z (the number of water molecules required to displace a protein from ligands) was derived. The in-tercept of this equation, log I, contains a number of constants a number of constants tha relate to the affinity of protein to the ligands. There is a good linear relationship betwwen log k's and log [H2O] under different chromatographic conditions. Although Z and log I varied with pH, slope j 1.74, the logarithm of hte molar concentration of pure water. Hydrophobic interactions dominate the retention of proteins in HIC at high salt concentrations.

A new technology for renaturation with simultaneous purification of the recombinant human interferon- (rhIFN-) in downstream of biotechnology is presented. The strategies to develop the new technology in industry scale were suggested. Based on chemical equilibrium and molecular interactions, the principle of rhIFN-refolding by HPHIC was described. The kind of stationary and mobile phases were evaluated and found the former to contribute to the rhIFN-refolding more than the latter. The extract containing the rhIFN-in gram scale in 7.0mol L−1 guanidine hydrochloride solution of 700mL was directly pumped into a unit of simultaneous renaturation and purification of proteins (USRPP, 10×300mmi.d.) packed by small particle packings of hydrophobic interaction chromatography and a satisfactory recovery of bioactivity and mass of the rhIFN-was obtained. With flow rate 100mLmin−1 and a gradient elution by only one step in 4h, the purity and specific bioactivity approach to 95% and 8.7×107 IU−1mg, respectively. To evaluate the goodness of the presented new technology in this study, a usual method with the renaturation by dilution method firstly and then purification with a series of LC in literature was employed to compare with each other. The obtained result in terms of purity, recoveries of mass and bioactivity, cost time as well as expenses, the former is much better than the latter. Comparing the total bioactivity of rhIFN- in the extract before to that after the renaturation by the USRPP, the total bioactivity of rhIFN- increased 62-fold.

变性蛋白表面的疏水氯基酸残基有与疏水色谱固定相（STHIC）颗粒相互作用的倾向，两者之间的疏水相互怍用能够抑制变性蛋白分子间的相互聚集。同时疏水色谱固定相还能在分子水平上给变性蛋白分子提供足够高的能量，使其瞬时脱水并折叠成其天然构象或不同的折叠中间体。变性蛋白在疏水界面上的折叠不仅取决于其氨基酸之间的特异性相互作用及疏水色谱固定相的结构，而且还取决于固定相和流动相之间的协同作用。同时，还提出了高效疏水相互色谱（IIPHIC）进行蛋白折叠的机理及其进行蛋白折叠时能实现质量控制的原理。在适当的色谱条件下，HPHIC可使几种变性蛋白一步实现复性及同时纯化。此外，还设计制造出了直径比柱长大得多的实验室型和制备型“变性蛋白复性及同时纯化装置，LISRPP”，该“装置”具有完全除去变性剂、使蛋白质复性，与杂蛋白分离及易于回收变性剂的“一石四鸟“功能。该“装置”对变性蛋白的复性和纯化效率与通常使用的长柱相当。在制备规模情况下，该“装置”可以在低压梯度条件下简便、快速、而经济地应用于重组蛋白药物的制备。文中以重组人干扰素。Y为例。说明了制备型“装置”在其复性及同时纯化生产工艺中的应用。

The monodisperse, poly (glycidylmethacrylate-co-ethylenedimethacrylate) beads with macroporous in the range of 8.0-12.0ml were prepared by a single-step swelling and polymerization method. The seed particles prepared by dispersion polymerization exhibited good absorption of the monomer phase. The pore size distribution of the beads was evaluated by gel permeation chromatography and mercury instrusion method. Based on this media, a hydrophobic interaction chromatographic (HIC) stationary phase for HPLC was synthesized by a new chemically modified method. The prepared resin has advantages for biopolymer separation, high column efficiency, low column backpressure, high protein mass recovery and good resolution for proteins. The dynamic protein loading capacity of the synthesized HIC packings was 40.0mg/ml. Six proteins were fast separated in less than 8.0 min using the synthesized HIC stationary phase. The HIC resin was firstly used for the purification and simultaneous renaturation of recombinant human interferon (rhIFN-) in the extract solution containing 7.0mol/l guanidine hydrochloride with only one step. The purity and specific bioactivity of the purified of rhIFN was found more than 95% and 1.3

本支依据溶液中溶质溶剜和吸附剂问相互作用的五个热力学平街，推导出了一个适用于多种吸附类型的吸附剂、溶质和溶剂的计量置换吸附模型。其核心是：溶质分子被吸附剂吸附时必然要释放出一定计量曲溶剂分子。用文献中已发表的原始数学对该甓型进行了挂验，并用奉模型的线性参数值对液一固厦咐现象进行了定量曲阐述。在Langmuir模型做了多方面比较后，发现以本模型为优。