Preparaton Of New Thiophilic Chromatography Monolithic Column And Application

Monolithic materials have received comprehensive attention and have been applied widely in various fields.Because they have the advantages of easy preparation,high mechanical strength,and low cost,etc.It also be called the fourth generation of separation matrix.Thiophilic chromatography is a powerful and simple method for the purification of proteins-in particular,immunoglobulins.It relies upon specific interaction between the immunoglobulin of thioether group and thiophilic ligand of sulfone group.However,at present,the main material of thiophilic chromatography materials have disadvantages of low mechanical strength,high cost,prone to loss of functional groups,and poor reproducibility.Hence,in this paper,thiophilic chromatography monolith was preparated for the first time.It was utilized mechanism of the thiophilic chromatography and advantages of the monolithic material.Besides,we also studied that the thiophilic chromatography column material for the enrichment and separation of small organic molecular containing disulfides,using the mechanism of thiophilic chromatography.To solve above issues,we carried out research from the following aspects:1.A novel thiophilic chromatography monolith was designed and synthesized by the method of post-modification.Firstly,The poly(MAA-co-MBAA)monolith was prepared from mixtures consisting of N,N-methylenebisacrylamide(MBAA)and glycidyl methacrylate(GMA)are dissoved in dodecanol and dimethyl sulfoxide(DMSO).The epoxy group of poly(MAA-co-MBAA)monolith was functionalized by sulfanilamide(SAA)to provide the sulphone group that necessary for SAA-poly(GMA-co-MBAA)thiophilic chromatography monolith.The monolith showed several significant advantages,including low cost,crosslink skeleton,good stability.This work presented potentially valuable of the novel thiophilic capillary monolithic columns for the antibody IgY enrichment and separation.2.A novel and facile one-step copolymerization approach was developed for the first preparation of well-controlled 3D skeletal thiophilic chromatography monolith.The poly(TEPIC-co-SAA)thiophilic chromatography monolith was prepared via ring-opening polymerization.The monolith set tris(2,3-epoxypropyl)isocyanurate(TEPIC)as a crosslinking agent,and sulfanilamide(SAA)as the functional monomer which provide the sulphone group,respectively.The obtained poly(TEPIC-co-SAA)thiophilic chromatography monolith showed the advantages of high mechanical and chemical stabilites.Benefiting from the step-growth polymerization process,the thiophilic monoliths possessed well-defined 3D framework microstructure.This mean that the pore size and the surface area of poly(TEPIC-co-SAA)thiophilic chromatography monoliths were controlled by adjusting porogen/monomer quality,porogen types and polymerization temperature.The porogen types was found as the most effective factor controlling the porous properties of poly(TEPIC-co-SAA)thiophilic chromatography monolith.The monolith was evaluated high selectivity for small organic molecular containing disulfides,such as 4,4'-Disulfanediyl diphenol and egg yolk immunoglobulin in capillary electrophoresis.This work presented potentially valuable of the novel thiophilic monolith for the antibody IgY enrichment and separation.3.A novel and facile one-step copolymerization approach thiophilic monolith was preparation.The poly(TEPIC-co-SFA)thiophilic chromatography monolith was prepared via ring-opening polymerization.The thiophilic monolith set TEPIC as a crosslinking agent,and sulfamide(SFA)as the functional monomer which provide the sulphone group,respectively.The obtained poly(TEPIC-co-SFA)thiophilic chromatography monolith showed the advantages of high mechanical and chemical stabilites.Benefiting from the step-growth polymerization process,the thiophilic monoliths possessed well-defined 3D framework microstructure.The thiophilic chromatography monolith was evaluatedhigh selectivity for small organic molecular containing disulfides,such as4,4'-Disulfanediyl diphenol,3H-1,2-benzene and sulfonated phenol2,3-ketone and egg yolk immunoglobulin in capillary electrophoresis.This work presented potentially valuable of the novel thiophilic monolithic columns for the organic disulfides enrichment and separation.