Investigations On The Separation And Purification Of Hemoglobin By Use Of Metal-organic Frameworks Y?BTC??H₂O?₆

Proteins are the basis of living bodies and important part of biological cells,which own close relationships to various life activities.As a kind of biological macromolecules with bioactivities,proteins generally have poor stabilities and tend to undergo conformational change in the process of purification,therefore,the development of mild and efficient extraction methods for the isolation of protein from complex sample matrixes becomes a key issue in proteomics and protein engineering.Metal-organic frameworks(MOFs)are a kind of coordination compounds,which usually have a metal ions or metal clusters as the coordination center,and oxygen or nitrogen containing porous organic ligands in the frameworks.In the present work,new organic frameworks was synthesized at room temperature using a rare earth metal Y and benzene-1,3,5-tricarboxylic acid,the production organic compound Y(BTC)(H2O)6 was further characterized by FT-IR,BET,XRD,SEM and surface charge analysis,and its application in proteins separation and purification was investigated.The performance of the obtained Y(BTC)(H2O)6 as adsorbent in the isolation of different proteins has been investigated.The experimental results indicated that Y(BTC)(H20)6 exhibits favorable selectivity toward the adsorption of neutral proteins.With hemoglobin as a model,the adsorption performance of Y(BTC)(H2O)6 is thoroughly studied.At pH 6.0,the adsorption efficiency of 87.7%for 100 ?g mL-1 hemoglobin in 1 mL sample solution was obtained when 0.40 mg of Y(BTC)(H2O)6 was employed.The adsorption behavior of hemoglobin onto the Y(BTC)(H2O)6 compound is consistent with Langmuir adsorption model,corresponding to a theoretical adsorption capacity of 555.6 mg g-1.The retained hemoglobin could be recovered by using 0.5%(m/v)SDS solution as the stripping reagent,the recovery of 77.3%was obtained.Circular dichroism spectra show no obviously difference on the conformation of hemoglobin after revovered by SDS.A solid-phase extraction protocol is developed for the isolation of hemoglobin from human blood and sodium dodecyl sulfate polyacrylamide gel electrophoresis was used to validate the purifity and separation.It indicated that the Y(BTC)(H2O)6 MOFs pose good ability for proteins separation and purification in complex matrix.