蛋白质在疏水性相互作用色谱介质上吸附的量热学研究-生物化工专业论文.docxVIP

ABSTRACT Hydrophobic interaction chromatography (HIC)， as one of important techniques in downstream bioprocessi吨， has been extensively applied in the pu盯1Ir汀讯i biological macromoωle町c时ud创es and nanopar川ticles队， such as proteins. The experimental evidences indicated that protein adsorption on HIC adsorbent may give rise to a loss of protein activity and conformational changes. In the current research ，the f conformational change of BSA on HIC adsorbents，Phenyl S叩harose 6 Fast Flow series，w制 investigated by calorimetrical analysis on a basis of isothermal titration calorimeter and van t Hoff method. FirstIy，the static adsorption of BSA on Phenyl Sepharose 6 FF was investigated at different conditions to assess the effect of salt types and concentration ， ligand density，temperature on the adsorption equilibria.ηle results showed that a stronger adsorption of BAS on HIC adsorbents could occur in presence of sodium sulfate compared with that in ammonium sulfate. 丁he protein adsorption w邸 more promoted by increasing sah concentration and ligand density. At a temperature range of 180C to 250C，the adsorption of BSA on Phenyl Sepharose 6 FF was an adsorption-dominated process while it transformed to a partitiondominated process at higher temperatures over 250C. Secondly，calorimetry of BSA adsorption on the HIC adsorbent was analyzed by l丁C，and then the Gibbs 仕ee ene咱y change e.G，enthalpy !:Jl， entropy change e.S and heat capacity change e.Cp ωuld be calculated from thermodynamic data collected in ITC experiments. 丁he result showed that adsorption of BSA on HIC adsorbent was typical of enthalpy-entropy compensation (巴巴C). During protein adsorption ，the unfavorable entropy change was compensated by a part of favorable enthalpy change and thus adsorption was driven by pa此ial enthalpy change. In a stronger adsorption of protein on HIC adsorbent ，the EEC became to weaken. During the adsorption process， moreove飞 adsorbed BSA on H